Immunology of Herpes Simplex Virus Infection: …...[CANCER RESEARCH 36, 836-844, February 1976] Immunology of Herpes Simplex Virus Infection: Relevance to Herpes Simplex Virus Vaccines

[CANCER RESEARCH 36, 836-844, February 1976]

Immunology of Herpes Simplex Virus Infection: Relevance to Herpes Simplex Virus Vaccines and Cervical Cancer

A n d r e J . N a h m i a s / S t e v e n L. S h o r e , S t e v e K o h l , S t u a r t E. S t a r r , a n d R o b e r t B. A s h m a n

Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30303 [A. J. N., S, K., S. E. S., R. B. A. ], and ~/irology Division, Center for Disease Control, USPHS Atlanta, Georgia 30322 [S. L. S.]

S u m m a r y

The immunology of herpes simplex infections has been reviewed, part icularly in relation to potential herpes simplex virus (HSV) vaccines, and to the association between HSV and cervical cancer. Relevant data from humans, experimental animals, and in v i t r o systems implicate both specific immune mechanisms and nonspecif ic factors in the course of HSV infections. There appear to be complex interactions between the various populat ions of mononuclear cells, macrophages, and lymphokines or other humoral factors. It is not yet possible, however, to pinpoint the crucial factors determining the different manifestat ions of the viral infection (primary infection, endogenous recurrence, or exogenous reinfection) in the various human hosts, although genetic factors may be important. While numerous animal models of HSV infection are available for the evaluation of HSV vaccines, models of HSV-induced cervical cancer re- quire further exploration.

I n t r o d u c t i o n

The past 15 years have witnessed a gradual, then explo- sive, transit ion from the purely serological to the more comprehensive immunolog ica l aspects of HSV-13 and HSV- 2 infections. The st imuli for improved understanding of host immune mechanisms to these viruses have come from several direct ions. Among these are (a) the large variety of c l in icopathological manifestat ions depending on the type of host, part icular ly the more severe disease observed in the immunosuppressed individual and in the newborn; (b) the possible role of immune factors in the reactivation or control of herpetic recurrences and the potential use of im- munomodulators for the prevention of such recurrences; (c) the association of HSV with human cancers; and (d) the development of HSV vaccines.

Coincidental ly, in recent years it has become appreciated that immune mechanisms may operate, not only on the

' Presented at the symposium "Immunological Control of Virus-associated Tumors in Man: Prospects and Problems," April 7 to 9, 1975, Bethesda, Md. Supported by Contract 4-3393 from the Virus Cancer Program, National Cancer Institute, and Grant DE 03924 from the National Institutes of Dental Research, NIH, USPHS.

= Presenter. a The abbreviations used are: HSV-1, herpes simplex virus type 1, HSV-2,

herpes simplex virus type 2; HSV, herpes simplex virus; CSF, cerebrospinal fluid; MIF, macrophage inhibition factor; i.c., intracerebral; BCG, Bacillus Calmette-Guerin; i.g., intragenital; CIS, carcinoma in situ.

virus itself, but also on virus-infected cells that carry HSV- specific antigens on their surfaces. This has, in turn, ex- panded the number of assays that can be used to measure humoral and cellular immune mechanisms and their interactions. Al though immune responses to HSV may well differ, depending on the type or subtype and whether fresh iso- lates or laboratory-passaged strains are involved, we be- lieve that restricting the discussion to HSV-2 strains alone would leave out potential ly important observations made with regard to other strains. This report reviews current information concerning the immunology of HSV infections in humans. Then fol lows a presentation of studies relevant to the mechanist ic aspects of the various immunological modalit ies and their interrelationships in animals and in v i t ro . We conclude with a discussion of certain immunological aspects related to cervical cancer and to HSV vaccines.

I m m u n o l o g y of H S V I n f e c t i o n in H u m a n s

Two lines of approach have been fol lowed in efforts to unravel immune mechanisms that might be operative in humans. The first has been to categorize special hosts who are susceptible to unusually severe HSV infections in an attempt to identify the critical parameters of the immune response. The more recent approach has been to apply a variety of immunologica l assays to individuals with various types of HSV infection in order to obtain direct information on immune funct ion. Because there may be variations in the immune responses associated with infection in special hosts with different types of HSV infection (primary or recurrent), 4 relevant informat ion is discussed below in its clinical context.

T h e I m m u n o s u p p r e s s e d or I m m u n o d e f i c i e n t Host . It is still unclear whether HSV infections are reactivated with greater frequency in the immunosuppressed host, as is the case with cytomegalovirus or varicella-zoster virus infections. What is clear, however, is that the recrudescences 4 of HSV-1 and/or HSV-2 tend to be more severe and chronic in immunological ly compromised individuals. The immuno- suppressive agents incr iminated include azothioprine, corti- costeroids, and ant i lymphocyte serum (9, 50, 54, 71, 89). Severe local HSV infections are also more common in individuals with cancers, particularly of the lymphoid organs (46, 56). Viral disseminat ion to visceral organs is infrequent

4 Recurrent infections are defined here as those occurring with or without clinical manifestations; recrudescent infections are limited to those with clinical manifestations.

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Immunology of HSV Infection

in these kinds of patients, but is more commonly observed in individuals with extensive burns (22) and in severely malnourished children, particularly in association with measles (4, 98).

Although several defects in immune responses can be found in these clinical conditions, a common theme is their association with defects in cellular immunity. Defects in cellular immunity are also found in children with the Wis- kott-Aldrich syndrome and individuals with circulating lym- phocytotoxic antibodies, some of whom develop the more severe forms of herpetic infection (35, 94). In contrast, severe HSV infections appear to be rare in individuals with defects in humoral immunity. In 2 cases of viral encephalitis in children with congenital X-linked hypogammaglobuline- mia, HSV was recovered from the brain at autopsy (43). However, the administration of attenuated measles virus vaccine a few days before the onset of symptoms in one child and the concomitant recovery of an enterovirus from the brain of the other child, suggest that viral-associated depression of cellular immunity might have contributed to the development of HSV encephalitis. An active effect of low levels of HSV antibodies in patients with congenital or acquired immunoglobul in deficiencies cannot be ruled out completely, because low titers of antibodies to other viruses can be detected in the serum of such patients.

Newborns. The contrast between the high frequency of disseminated HSV (most usually HSV-2) infection which occurs in the newborn and the rarity of viral dissemination in older individuals is consistent with the interpretation that the maturity of the immune system plays an important role in host resistance to the virus (61). However, any such defect in the newborn must be short-lived, since viral dissemination to visceral organs rarely occurs after 3 weeks of age.

Although transplacentally acquired HSV antibodies are not fully protective to the newborn, infection in such cases tends to be localized to the skin, eyes, or central nervous system, without evidence of viral dissemination to visceral organs (61, 66). It is still undetermined whether cases of neonatal herpes occurring in the face of transplacental antibodies are the exception rather than the rule.

We have recently demonstrated that cord blood mononuclear cells of normal newborns have the capacity to act synergistically with HSV antibodies in an antibody-dependent lymphocyte-mediated cytotoxicity assay against HSV-1- or HSV-2-infected target cells (87). Although, compared with adult cells tested at the same effector:target cell ratio, newborn mononuclear cells showed slightly lower cytotoxicity, the total cytotoxicity of cord blood was comparable when adjusted for the higher content of mononuclear cells in newborns. Studies on pairs of maternal and cord blood sera demonstrated the transfer of the lymphocyte-dependent antibody in equivalent titers, providing further evidence for the IgG nature of this antibody.

Infected newborns have been found to produce IgM specific for HSV within 1 to 3 weeks after onset of infection (66). These antibodies continue to increase during the 1st 2 months and may be detectable for as long as 1 year. Some of the IgM antibodies in the serum of infected infants are directed against allotypic determinants on the maternal IgG.

The lymphocytes of some infected newborns have been found to respond to HSV antigens in vitro several weeks after onset of the infection (91). The temporal sequence of the development of cell-mediated responses and various lymphokines in these newborns requires concerted study; such studies are limited by the relatively low survival rate of these patients in any one institution.

Infection of the Nervous System. In individuals other than newborns, most cases of HSV encephalitis are caused by HSV-1; cases of meningitis are related to HSV-2 (12). HSV-2 can be isolated from the buffy coat of peripheral blood and from the CSF in cases of meningitis, suggesting the potential of HSV-2 to disseminate via the blood. It is still puzzling why HSV encephalit is occurs in apparently normal individuals, particularly in those with evidence of HSV infection before the neurological disease develops. The 2 cases of HSV encephalitis in agammaglobulinemics with concomitant viral infections (43), which might have suppressed cellular immune responses, indicate the need for more intensive investigations along these lines. HSV encephalitis, however, has not been observed with any greater frequency in immunosuppressed or immunodeficient individuals. In contrast, subacute HSV encephalitis has been observed in an anergic patient (72).

High levels of interferon have been detected in the blood and spinal fluid of a severely affected 4-month-old infant with HSV encephalit is (5). The source of the interferon and its role in the disease process are unknown. HSV antigens have been detected by immunofluorescent techniques in CSF leukocytes from patients with encephalitis, although infectious virus could not be isolated from that body fluid (15). HSV antibodies can be measured in the CSF of individuals with HSV encephalit is, although they are demonstrable relatively late after onset (40). The higher titer of HSV antibodies in the CSF, compared with that found in the serum, implies local synthesis of antibodies within the nervous system.

Primary HSV Infection. This is defined as an HSV-1 or HSV-2 infection occurr ing in an individual with no prior infection with either HSV type. There are no data to explain why primary HSV-1 and HSV-2 infections are subclinical in most individuals, yet can be very severe in a few others. Immunogenetic studies would be particularly relevant to this point. The more severe manifestations of primary HSV-2 infection during pregnancy (32) suggest that hormonal factors and/or the relative immunosuppression occurring during pregnancy may be involved.

Serological studies on primary HSV-1 and HSV-2 infections have revealed that tgM antibodies are the first to be demonstrated, usually followed by the development of IgG and IgA antibodies (24, 60). Lymphocyte-dependent antibodies appear around the time IgG antibodies are first detected (86). Complement-dependent neutralizing antibodies can be detected earlier in the serum of individuals with primary HSV infections (112).

Assays for cell-mediated responses have been applied in a very limited number of individuals with primary HSV-1 and HSV-2 infections. Lymphocyte transformation to HSV antigens has been demonstrated between the 2rid and 4th week after onset of infection (91).

Recurrent HSV Infection. HSV-2 infection occurs more

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commonly in individuals who have had pr ior HSV-1 infec- t ion than as a primary infection. The cl inical manifestations tend to be mi lder in such cases. Immunological studies of recurrent HSV-2 infection are rendered more complex by several problems: (a) def ining individuals with pure HSV-2 recurrences, i .e., without prior HSV-1 exposure; (b) the possibi l i ty of exogenous reinfection with the same or different subtype of HSV-2; and (c) the di f f icul ty of ensuring the absence of subcl inical recurrences, when sequential studies are performed.

The mechanisms of viral persistence after a primary infec- t ion are as yet unknown, but it appears that the viral genome lies dormant in the tr igeminal gangl ia in patients with labial or ocular infections (3) and in the sacral ganglia in patients with genital herpes (2). The various tr iggering mechanisms, which include sunshine, menstruation, sexual intercourse, fever, and emotional stress, provide little information about the underly ing pathobiology.

Patients with recurrent HSV infections usually possess high levels of serum antibody. Serum titers of 1:100,000 can be detected with an ant ibody-dependent lymphocyte cytotoxici ty assay (86). In most patients there is no change in ant ibody titers, as measured by neutral ization tests, with or wi thout the addit ion of complement (27), or by complement- f ixation assays, fo l lowing a recurrence. However, with the use of more sensitive serological tests, such as radioimmu- noassay and immunof luorescence, rises in titers of IgM, IgG, or IgA antibodies can be demonstrated in some individuals after a recrudescence (60). In contrast to an earlier report (101), other workers have shown that levels of IgA in oral or ocular secretions are not depressed in recurrent herpes, and that the titers of HSV antibodies in secretions do not f luctuate with clinical status (8, 17).

Most recent work has centered around the cellular immune responses to herpetic infection, albeit, conducted most often in individuals with recurrent HSV-1 infection. When skin testing with HSV antigens was performed, no dif ferences were found in the response of patients with recrudescences, compared with that of seropositive, asymp- tomat ic individuals (6, 82, 110). Skin test reactivity corre- lated well with the presence of c irculat ing HSV antibodies.

Peripheral blood lymphocytes from seroposit ive individuals transform when exposed to HSV antigens in vitro (74, 78, 81, 91, 109). In general, responses are higher with the homologous HSV type. Confl ict ing results have been obtained as regards the changes in the lymphocyte st imulat ion index induced by HSV antigens in relation to recrudescences. Some studies have shown no apparent differences in the lymphocyte stimulation index at different times during or after the herpetic recrudescence (74,109). Other studies have shown an increase in the st imulat ion index shortly after the recrudescence, and a fall thereafter (78). Varia- t ions in the antigens used by the various investigators might explain some of the differences observed. Lymphocyte responses to phytohemagglut in in, purif ied protein derivative, streptokinase, vaccinia, and Candida a lb icans were normal in both seroposit ive patients wi thout recrudescences and in patients with recrudescence (109).

The role of lymphokines in recurrent herpes is unclear as yet because of seemingly conf l ict ing data. Lymphocytes

from patients with recrudescences, when st imulated with HSV antigens, produce less MIF than lymphocytes from seropositive individuals wi thout recrudescences (109). However, product ion of leukocyte chemotact ic factor and lymphotoxin were not found to differ between the 2 groups (78). A sequential study of interferon product ion by HSV- stimulated leukocytes, in patients with recrudescences, showed that maximum levels were attained 2 to 6 weeks after onset of the recrudescence (74). The leukocytes pro- ducing the interferon have been identified as being T-cells (102). Interferon induced by HSV, using combined macrophage lymphocyte cultures, has been found to differ in its pH and temperature labil ity from human interferon induced in other systems (103). Individuals with the highest levels of leukocyte-produced interferon after a recrudescence tended to have a longer duration between recrudescences. A similar pattern was noted when leukcoyte migration inhibitory factor was measured (69).

Direct lymphocyte cytotoxici ty has been reported against cells acutely or chronical ly infected with HSV-I. With the acutely infected cells, lymphocytes from patients with recurrences taken during the quiescent period were specifically cytotoxic (83). With the chronical ly infected target cells, lymphocyte cytotoxic responses of patients wi th recrudescences (during the quiescent phase) were signif icantly lower than those of seropositive individuals wi th no history of recurrences (100). Ant ibody-dependent lymphocyte-mediated cytotoxici ty might be confused with direct lymphocyte-mediated cytotoxici ty if HSV antibody remains on the membranes of the effector cells after the lymphocytes are washed.

Susceptibi l i ty to recurrent herpetic infection may also be associated with genetic factors. Russell and Schlant (84) recently demonstrated a higher frequency of HLA-1 transplantation antigens in patients with frequent herpetic recrudescences than in a control population.

Erythema Multiforme. It is cl inically appreciated that individuals with HSV infections may demonstrate the allergic manifestations of erythema mult i forme (85, 95). The immu- nopathological mechanisms are unclear, al though they are likely to be related to the formation of ant igen-ant ibody complexes. The allergic skin manifestation can be induced by the administrat ion of inactivated HSV preparations (85).

Immunology of HSV in Experimental Animals

Immunological studies have been conducted in a variety of experimental animals, particularly in mice, guinea pigs, and rabbits. The fo l lowing factors have been implicated in host responses to HSV infection: genetics, temperature, age, and humoral and cellular immune responses.

Genetics. A recent report suggests that genetic factors are operational in the natural resistance of mice to i.p. inoculat ion with a newly isolated HSV-1 (48). The 11 strains of mice could be categorized as resistant, moderately susceptible, and very susceptible. Fj crosses indicated that resistance is governed by a dominant gene, al though it is apparently not l inked with a histocompatibi l i ty locus.

Temperature. Adult mice, inoculated i.c. with HSV-1 or




i.p. with HSV-2, experience signif icantly lower mortal i ty when housed at 34-36 ~ than when housed at ambient temperature (22o) 3 (49). Although symptomatology was probably modif ied by metabolic changes, the decreased mortal i ty may have been a consequence of temperature-dependent inhibit ion of viral replication. Thermal inhibi t ion of HSV replication has been reported in tissue culture systems (13), HSV-2 usually being more temperature labile than HSV-I.

Age. Newborn mice have repeatedly been shown to be more susceptible to extraneural HSV inoculat ion than are adult mice. Johnson (31) has demonstrated that a barrier to the spread of virus inoculated extraneural ly developed with age and that the inhibit ion of viral disseminat ion depended on the peritoneal and tissue macrophages. Other workers (28) found that syngeneic peritoneal macrophages from adult mice protected neonatal mice from i.p. challenge with HSV. In HSV-infected older mice, specif ic inhibi t ion of macrophage funct ion by the administrat ion of silica particles or ant imacrophage serum led to viral disseminat ion and early death (113). Proteose peptone-st imulated macrophages were more eff icient in conferr ing protect ion against HSV infection than unstimulated cells; the enhanced resistance was associated with more eff icient phagocytosis and greater product ion of interferon. In contrast, macrophages from neonatal mice did not respond to proteose peptone stimulation (28). These f indings imply that macrophage im- maturity is an important factor in the increased susceptibil- ity of neonatal animals to viral disseminat ion.

BCG has been shown to increase the resistance of newborn mice to subsequent i.p. chal lenge with HSV-2, probably by activating macrophages. '~ BCG also decreased the mortality of rabbits after corneal or intravaginal inoculat ion with HSV-2 (36). However, BCG had no protective effect against intravaginal inoculat ion of HSV-2 in adult mice, except when combined with HSV-2 antiserum (1). Levami- zole has been found to confer some degree of protection to newborn rats inoculated with HSV-2 (21).

Humoral Immune Responses. HSV can be recovered from the spinal ganglia of previously infected mice in which latency had been established (10, 93,105). Active immunization with HSV-2 does not affect the establ ishment of latent gangl ionic infection, whereas immunizat ion with HSV-1 con- fers a moderate degree of protection (105). When ganglia latently infected with HSV-1 were transplanted to nonin- fected syngeneic mice, both infectious virus and viral antigens could be demonstrated in the recipient animals. Less virus was expressed in ganglia transplanted to latently infected mice or to mice that had been passively immunized with anti-HSV IgG (93). Al though a simi lar phenomenon may be operative in suppressing the reactivation of virus from human ganglia, it is obviously not completely effective in inhibit ing recurrences in man.

Serum or ~,-globulin containing HSV antibodies can prevent infection if administered before, or within hours after, inoculation of virus (25, 26). Passive immunizat ion thereafter is not usually effective. HSV serum antibody can also act

s. Starr, A. Visintine, M. Tomeh, and A. Nahmias. Non-specific Immuno- therapy in Herpes Simplex Virus Type 2 Infection of Newborn Mice, submit- ted for publication.

Immunology of HSV In fec t ion

synergistically with peritoneal cells from non immune m ice to destroy herpes-infected cells in vitro (73). Immun iza t ion with inactivated HSV will induce a variable degree of p ro tec- t ion, depending on the route of virus challenge, but t he mechanisms of protect ion have not been fully clarif ied (25, 105). Nevertheless, mice or cebus monkeys genital ly infected with HSV-2 could be reinfected with the same v i rus by the genital route several months later (47).

The physical state of the virus and the virus dose p robab l y inf luence the type of immune response. Thus, rabbits i nocu - lated with infectious HSV developed both cel lular and hu - moral immunity, whereas those inoculated with UV- inact i - vated virus developed a predominant ly cellular response, with little or no neutral iz ing ant ibody (77). Inoculat ion w i t h an HSV-anti-HSV complex did not el icit either a cel lular or a humoral response. The type of assay used to measure an t i - bodies in rabbits fo l lowing HSV inoculat ion also appears to be important. Thus, the 1st ant ibody to appear is best measured with a complement- requi r ing neutral iz ing assay (41, 111).

Cel l -mediated Responses. The development of de layed hypersensitivity is acknowledged to be a cel l-mediated immune phenomenon. Guinea pigs sensitized to HSV d e m o n - strate a cutaneous response to viral antigens (38, 75). The t iming of the maximal response, the mononuclear nature o f the infiltrate, and the abil i ty to transfer reactivity with ce l l s but not with serum, have conf i rmed that these skin reac- t ions are those of classical delayed hypersensit ivity. Re la ted studies have shown that hypersensit ivity to HSV an t igens can also be induced in the cornea of both guinea pigs and rabbits (52, 53).

The importance of lymphocytes in the immune response has been conf i rmed by adoptive transfer exper iments and i n studies with immunosuppressed animals. Spleen cells f r o m HSV-l- immunized mice were shown to confer partial p ro tec - t ion to recipient mice chal lenged with the virus (19). Neo- natal thymectomy in mice depressed their resistance to HSV infection, al though neutral iz ing HSV antibodies were p ro - duced to almost the same extent in neonatally t hymec to - mized mice as in intact controls (55). HSV-infected m i ce treated with ant i lymphocyte serum showed an increase in the magnitude and durat ion of the viremia and g rea te r dissemination to the central nervous system (113). M i ce treated with ant i thymocyte serum showed different pa t te rns of survival, depending on the route of viral inoculat ion (62). Survival was signi f icant ly poorer in the ant i thymocyte serum-treated mice when they were chal lenged either i.p. o r i.g. However, in mice chal lenged i.c., increased survival and prolongat ion of survival t ime were observed, compared w i t h untreated controls.

The specif icity of the cel lular immune response to HSV has been demonstrated in rabbits by prior inoculat ion w i t h HSV-1 or HSV-2, fo l lowed by st imulat ion of splenic l y m p h o - cytes with the appropr iate antigens in vitro (79). L y m p h o - cytes responded more vigorously to the sensit izing than to the heterologous virus type. It was further shown that l ym- phocyte sensit ization occurred within 3 days after in fec t ion , with a peak response at 7 days, and that sensit ized ce l ls could still be detected up to 120 days after virus i nocu la t i on (76). In vitro, specif ical ly sensitized lymphocytes could be

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stimulated by virus alone and by virus bound to cells, but the reaction was abrogated if the cel l -bound virus was incu- bated with antiviral ant ibody.

The temporal relation of cellular and ant ibody-mediated immune responses has also been studied in experimental herpetic keratitis in rabbits (51). Lymphocyte cytotoxici ty and MIF were detected within the 1st 7 to 11 days postinocu- lation. Detection of neutral izing antibodies peaked at days 11 to 21 and that of complement-dependent cytotoxic antibodies peaked on Day 16. The authors concluded from these observations that the control of ocular HSV infection involves an early inf lammatory phase with macrophage reactivity and elaborat ion of MIF by sensitized lymphocytes. Transient virus-specif ic T-lymphocytes with effector reactivity, as well as neutrophi ls with chemotact ic activity, occur dur ing the stage of stromal keratitis. Ant ibody-dependent complement-mediated lysis later provides another phase of restriction of the infection.

HSV is susceptible to interferon and is interferogenic (23, 80, 96), but whether all strains are equally effective interferon inducers is not yet clear. Interferon was detected in the skin of mice 24 hr after the s.c. injection of HSV, and it increased to a peak 5 to 7 days later (96). Administrat ion of interferon inducers was effective in protect ing mice only when very small amounts of virus were used as inoculum (7). To date, no conclusive correlat ion between interferon levels and survival after HSV challenge has been demonstrated.

Virus-immune Interactions In Vitro

A fruitful area of investigation has been the examinat ion in vitro of HSV, virus-infected cells, and immune effectors. Such systems have facil i tated the dissection of the various specific and nonspecif ic effectors and their interrelationships. Since HSV can spread to other cells by extracel lular transmission or by direct cell-to-cell spread, it fol lows that immunolog ica l intervention can proceed at 3 levels: (a) uptake of virus by leukocytes, (b) neutral ization of extracellu- lar virus by humoral factors, and (c) destruct ion of virus- infected cells by humoral and cellular factors.

Uptake of Virus by Leukocytes. HSV is readily taken up by macrophages from both adult and neonatal mice (28, 31, 92). Viral antigens are detectable within 4 hr after infection in macrophages from adult mice, with maximal expression in 10 to 20% of the cells at 6 hr after infection. Infected macrophages from neonatal mice, in contrast to macrophages from adult mice, are able to infect surrounding cells. The titer of virus produced in suckl ing mouse macrophages is s igni f icant ly higher than that produced in adult mouse macrophages. The abil i ty of adult macrophages to l imit viral repl icat ion is not due to lack of viral adsorpt ion, because adult macrophages absorb 100 t imes more virus than do those of suckl ing mice (28). There is no block in synthesis of viral DNA or viral proteins, but, instead, a restriction of viral assembly (92). No data are yet available on the activity of macrophages in human HSV infection. It is tantal iz ing to infer from the mouse studies that a defect in macrophage funct ion might lead to visceral disseminat ion, such as occurs in newborns.

Whereas macrophages may help to restrict viral growth and dissemination, there is the contrasting f inding that peripheral blood lymphocytes may allow repl icat ion of HSV (59). Viral replication has been found to be facilitated in human lymphocytes stimulated with phytohemagglut in in, pokeweed, concanaval in A, and ant i lymphocyte serum (34, 59). The subpopulat ions of cells capable of sustaining viral replication are yet to be ascertained, as is the in vivo signifi- cance of these observations.

Effect of Humoral Factors. When moderate concentra- tions of HSV are exposed to specific ant ibody in vitro, a so- called "unneutra l izable" fraction remains (68, 104, 111). This can be rendered noninfect ious by adding either complement or ant i immunoglobul in . Several explanat ions have been offered for these observations: (a) an actual virolytic action of ant ibody and complement, as is seen in bacterial systems; (b) immunoaggregat ion of ant ibody-coated virus; and (c) an enhanced steric effect of ant iant ibody or complement in actually covering critical virus sites necessary for viral adsorption or penetration. The recent f ind ing (14) that the alternate complement pathway can also assist antibody in virus neutral ization may be of relevance to this issue.

The interaction of virus and immune lymphocytes has been shown to t r igger the release of migratory inhibitory factor, leukocyte inhibi tory factor, lymphotox in, and interferon (51, 69, 74, 78, 109). Ant igen-ant ibody complexes of HSV with IgG or IgM are also capable of s t imulat ing immune lymphocytes. Rabbit spleen lymphocytes exposed in vitro to HSV antigens demonstrate increased thymid ine uptake and interferon product ion if the donors are previously immunized with HSV (30, 76). Cells infected with virus and then challenged with HSV ant ibody and complement release a factor chemotatic for mononuclear and po lymorphonuclear leukocytes (97). HSV has also been found to depress human monocyte chemotaxis (33).

Effect on Virus-infected Cells. HSV-infected cells acquire virus-specific membrane antigens which al low recognit ion and subsequent attack by various immune effectors (58). Spleen cells from sensitized mice or guinea pigs are able to reduce the size of plaques formed after addit ion of a known number of HSV plaque-forming units (20, 88). The activity of the mouse spleen cells was not affected by removal of adherent cells, and no lymphotoxin was detectable in the culture supernatants. It has also been demonstrated that high ratios of casein-activated but nonimmune rabbit peritoneal exudate cells cause nonspecif ic cytotoxic i ty and prevent viral plaque format ion, although the addi t ion of HSV antibody and complement enhanced the reaction (44). With the HSV-1 strain and tissue culture used, complement-mediated ant ibody cytolysis was found to be active after cell- to-cell viral spread has already occurred. Leukocytes from rabbits immunized with either HSV-1 or complete Freund's adjuvant and st imulated with inactivated HSV-1 or purif ied protein derivative have been shown to inhibit viral replication in a plaque assay (45). In contrast to cytotoxic i ty assays, this effect could be demonstrated with effector-to- target cell ratios as low as 1:10. The supernatant fluid from these cultures was shown to contain interferon, which could also inhibit viral replication.

An observation that not only complicates the interpreta-




Immunology of HSV Infection

tion of these results, but also suggests an alternative mechanism for destroying virus-infected cells, is the demonstration that HSV-1 or HSV-2 antibody can act synergistically with immune or nonimmune mononuclear cells to induce cytolysis in both HSV-1- or HSV-2-infected target cells (83, 86, 87). The antibody, which is IgG, is effective at serum dilutions to 1:100,000, so that scrupulous washing of the mononuclear cells is necessary before all of this contaminat- ing antibody is removed. The human mononuclear cells have been partially characterized and appear to be neither T-cells, nor B-cells, nor monocytes; they are most probably K-cells. It has been suggested that the mechanism of target cell damage involves 2 stages: recognit ion of viral antigens on the infected cell by attachment of specific IgG, fol lowed by interaction of the effector cell with the Fc portion of the bound immunoglobul in molecule (86, 87).

A further complexity of the relationship between HSV and host immune factors is the finding of receptors for the Fc fragment of IgG on the surface of HSV infected cells and some of the HSV transformed hamster cells (107, 108). The importance of this observation is still unclear. However, the presence of the Fc receptor might protect the infected cell from antibody-mediated immune attack either by binding the Fc fragment of IgG antibody so that the Fc is unable to activate complement or cytotoxic effector cells, or by steri- cally inhibit ing the recognition of nearby virus-specific membrane antigen sites.

Notkin's group has attempted to synthesize the nonspecific and specific control of cell-to-cell spread by virus into a 2-step response encompassing specif ic recognition and nonspecific execution (44, 45). The response involves specific immunological recognition by previously sensitized leukocytes, antibody and complement, which react with virus or viral antigens on the surface of infected cells. This generates mediators from both complement and immune leukocytes which can attract inf lammatory cells to the site of infection. In addition, other mediators, such as lymphotoxin and interferon, are produced. The lymphokines and inf lammatory cells then act in a nonspecif ic fashion on infected and adjacent uninfected cells to prevent viral cell- to-cell spread.

Cervical Cancer

If HSV-2 is somehow causally related as an initiator or promoter of cervical carcinogenesis, it would be advanta- geous to use immunological means to abrogate either the initial infection or possibly the recurrent infection, which may be as important (65). Such an approach would have the added advantage of providing firm evidence for causality. It would therefore be desirable to understand, not only the immunology of the herpetic infections, but also that of cervical neoplasia, and their possible interactions.

There are several levels at which immune factors may affect the development and progression of cervical neoplasia: (a) rejection of the initially transformed cells, (b) prevention of the invasion of the basement membrane by the intraepithelial neoplasia, and (c) prevention of local exten- sion and distant metastasis of the invasive cancer.

The reversion of cervical dysplasia to normal in many women and the slow progression from dysplasia to CIS, and

from CIS to invasive cancer, might depend on immune factors which are depressed at critical periods. Coppelson and Reid (11) suggested that host immune factors might operate in this process, on the basis of their morphological studies. These workers detected mononuclear and phago- cytic cells in most histopathological sections of cervical dysplasia, CIS, and microinvasive cancer. Although leukocy- tic cells were usually found along the basement membrane, the foci were occasionally subepithelial and were sur- rounded by a basement membrane. The superficial epithe- lium itself was occasionally infiltrated with mononuclear cells, particularly around intraepithelial capillaries. Evi- dence of death of the cervical neoplastic cells was some- times associated with an intimate attachment of the mononuclear cells to the neoplastic cells and was most often observed in the stroma at the stage of CIS or microinvasion.

A limited number of studies have been directed specifically towards the immunological aspects of cervical neoplasia. Positive skin test reactions have been obtained with antigens prepared from the membranes of cervical cancer cells (29). In another study (67), skin testing with various antigens, such as Candida or streptokinase-streptodornase, showed a lower frequency of responses in women with the later stages of invasive cervical cancer, compared with those with intraepithelial neoplasia or early invasive cancer. A depression in in vitro lymphocyte transformation responses to HSV antigens has been demonstrated in patients with oral cancer, and increased st imulation in those with precancerous lesions (39). Antibodies to "tumor-specif ic antigens" of invasive cervical cancer tissues have been detected in the sera of women with cervical cancer by immunodif fusion techniques (42). Lymphocytes from women with cervical cancer have been reported to react against cultured cervical cancer cells in a microcytotoxici ty assay (16), and complement-dependent antibody cytotoxicity has also been demonstrated in a similar test system (106). An increase in titers of HSV-2-neutralizing antibody has been associated with a decrease in complement-dependent HSV antibodies with progression of cervical neoplasia (99).

Other immunological studies have used HSV-transformed hamster cells. Spleen cells from hamsters inoculated with HSV-1- or HSV-2-transformed cells, and from hamsters immunized with HSV-1, were specif ically cytotoxic for HSV-1- transformed cells in vitro (37). Sera from these animals did not cause complement-mediated antibody cytolysis of the HSV-transformed target cells. In another study (90), spleen lymphocytes from HSV-2 tumor-bearing animals were cytotoxic to both transformed and HSV-infected cells. Immune lymphocytes have also been used to detect the transfer of the HSV genome from transformed cells to HSV-susceptible cells (90).

Herpes Simplex Vaccines

The prospects and problems related to HSV vaccines will be discussed by other speakers in this Symposium. We would like to address ourselves to a few points with immunological overtones.

Of paramount importance are the various types of hosts to which an HSV-2 vaccine might be administered.

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T h e Ind iv idua l Not P r e v i o u s l y I n f e c t e d with HSV-1 or HSV-2 . In th is case, the vacc ine wou ld have to be admin- is tered a round the age of 6 mon ths to 1 year, the t ime of 1st exposu re to HSV- I . The possib le in f luence of t rans- p lacenta l an t i bod ies and the need for i m m u n i t y to pers ist in to ado lescence and adu l t life p resent s ign i f i can t di f f i - cul t ies.

T h e Ind iv idua l A l r e a d y I n f e c t e d with HSV- I . The HSV-2 vacc ine cou ld be given such ind iv idua ls before adoles- cence, p r io r to the usual 1st exposu re to HSV-2. Whe the r the vacc ine w o u l d p rov ide suf f ic ien t spec i f i c i m m u n i t y is p rob lema t i c , in v iew of the we l l - r ecogn i zed inabi l i ty of a p r i o r HSV-1 in fec t ion to p ro tec t aga ins t HSV-2 in fec t ion (63).

T h e Ind iv idua l A l r e a d y I n f e c t e d with HSV-2 . Here, the HSV-2 vacc ine m igh t be of po ten t ia l value in p revent ing recu r rences . S ince the recu r ren t in fec t ion m igh t be as im- po r tan t or even more i m p o r t a n t than the init ial herpet ic in fec t ion in caus ing neop las t i c t r a n s f o r m a t i o n s (65), such an a p p r o a c h cou ld p rove va luable.

These cons ide ra t i ons sugges t that , s ince there may wel l be spec i f i c responses to each of the t w o vi rus types, exp lo- ra t ion of HSV vacc ines shou ld inc lude not on ly HSV-2 but a lso HSV- I .

In con t ras t to the o the r human herpesv i ruses, we are fo r tuna te , w i th HSV, to have a p le tho ra of expe r imen ta l an ima l mode ls for gen i ta l or nongen i ta l HSV-1 or HSV-2 in fec t ions w h i c h can be used to eva luate p rospec t i ve vac- c ines. These inc lude n o n h u m a n pr imates , as wel l as rabbi ts and var ious roden ts (21, 36, 47, 52, 64). Fu r the rmore , a mouse model fo r de l i nea t i ng the ef fect of act ive immun iza - t ion on the la tency of HSV-1 and HSV-2 has recent ly been deve loped (105). Dose response, m o d e of adm in i s t r a t i on , types of i m m u n i t y e l ic i ted, and the du ra t i on of p ro tec t i on can readi ly be s tud ied in such systems. In con t ras t to the ready avai lab i l i ty of an ima l mode ls for HSV in fec t ion , a sa t i s fac to ry an ima l mode l for the eva lua t ion of the pro tec- t ive ef fect of HSV vacc ines aga inst cerv ical neop las ia has yet to be deve loped . Mode l systems that m igh t be cons id- ered inc lude Cebus m o n k e y s and mice. The ut i l i ty of the m o n k e y mode l awa i t s resul ts of a t tempts to p roduce cervical neoplas ia af ter gen i ta l i nocu la t i on of HSV-2 (70); however, p re l im ina ry resul ts in the mur ine expe r imen ta l system ind ica te that cerv ical neop las ia can be induced af ter geni ta l i nocu la t i on of HSV (57, 60). The es tab l i shmen t of a sat is fac- to ry an imal mode l is vital in v iew of the d e m o n s t r a t i o n tha t i m m u n i z a t i o n w i th HSV enhances t u m o r d e v e l o p m e n t in hamste rs i nocu la ted w i t h HSV- t rans fo rmed t u m o r s (18).

O the r cons ide ra t i ons inc lude not on ly po tent ia l reac t ino- gen ic side ef fects of any p rospec t i ve vacc ine, but also long- te rm i m m u n o p a t h o l o g i c a l ef fects. It has been s h o w n , fo r ins tance, that e r y thema mu l t i f o rme , w h i c h can o c c u r w i th recu r ren t HSV in fec t ion , can also be p rec ip i ta ted by admin - is t ra t ion of inac t iva ted HSV (85).

R e f e r e n c e s

1. Baker, M. B., Larson, C. L., Ushijima, R. N., and Anderson, F. D. Resistance of Female Mice to Vaginal Infection Induced by Herpesvirus Hominis Type 2: .Effects of Immunization with Mycobacterium bovis, Intravenous Injection of Specific Herpesvirus Hominis Type 2 Antiserum

and a Combination of These Procedures. Infect. Immunity, 10: 1230- 1234, 1974.

2. Baringer, J. R. Recovery of Herpes Simplex Virus from Human Sacral Ganglions. New Engl. J. Med., 291: 828-830, 1974.

3. Baringer, J. R., and Swoveland, P. Recovery of Herpes Simplex Virus from Human Trigeminal Ganglions. New Engl. J. Med., 288: 648-650, 1973.

4. Becker, W. B., Kipps, A., and McKenzie, D. Disseminated Herpes Sim- plex Virus Infection: Its Pathogenesis Based on Virological and Patho- logical Studies in 33 Cases. Am. J. Diseases Children, 115: 1-8, 1968.

5. Bellanti, J. A., Catalano, L. W., and Chambers, R. W. Herpes Simplex Encephalitis: Virologic and Serologic Study of a Patient Treated with an Interferon Inducer. Pediat. Pharmacol. Therap., 78: 136-145, 1971.

6. Bubola, D., and Olivetti, L. L'lntradermoreazione Con Virus Erpetico Innattivato. I. Analisi dei Jattori che Influenzano la Reasione. G. Ital. Dermatol., 109: 363-376, 1968.

7. Catalano, L. W., London, W. T., Rice, J. M., and Sever, J. L. Prophylac- tic and Therapeutic Use of Poty (I) Poty (C) [Poly-D-lysine] against Herpesvirus Encephalitis in Mice. Proc. Soc. Exptl. Biol. Med., 140: 66- 71, 1972.

8. Centifanto, Y. M., Little, J. M., and Kaufman, H. E. The Relationship between Virus Chemotherapy, Secretory Antibody Formation and Recur- rent Herpetic Disease. Ann. N. Y. Acad. Sci., 173: 649-656, 1970.

9. Chiba, S., Ikeda, S., Agatsuma, Y., Nakao, T., and Sabeki, Y. Active Infection with Cytomegalovirus and Herpes Group Viruses in Children Receiving Corticosteroid Therapy. Tohoku J. Exptl. Med., 106: 265-270, 1972.

10. Cook, M. L., Bastone, V. B., and Stevens, J. G. Evidence That New- borns Harbour Latent Herpes Simplex Virus. Infect. Immunity, 9: 946- 951, 1974.

11. Coppleson, M., and Reid, B. Preclinical Carcinoma of the Cervix Uteri. Oxford, England: Pergamon Press, Inc,, 1967.

12. Craig, C. P., and Nahmias, A. J. Different Patterns of Neurologic Involve- ment with Herpes Simplex Virus Types 1 and 2: Isolation of Herpes Simplex Virus Type 2 from the Buffy Coat of Two Adults with Meningitis. J. Infect. Diseases, 127: 365-373, 1973.

13. Crouch, N. A., and Rapp, F. Cell-dependent Differences in the Produc- tion of Infectious Herpes Simplex Virus at a Supraoptimal Temperature. J. Virol., 9: 223-230, 1972.

14. Daniels, C. A., Sandberg, A. L., and Notkins, A. L. Neutralization of Herpes Simplex Virus by the Alternate Complement Pathway. Federa- tion Proc., 34: 853, 1975.

15. Dayan, A. D., and Stokes, M. I. Rapid Diagnosis of Encephalitis by Immunofluorescent Examination of Cerebrospinal Fluid Cells. Lancet, 1:177-179, 1973.

16. DiSaia, P., Sinkovics, J. G., Rutledge, F. N., and Smith, J. P. Cell- mediated Immunity to Human Malignant Cells. Am. J. Obstet. Gynecol. 114: 979-984, 1972.

17. Douglas, R. G., Jr., and Couch, R. B. A Prospective Study of Chronic Herpes Simplex Infection and Recurrent Herpes Labialis in Humans. J. Immunol., 104: 289-295, 1970.

18. Duff, R., Doller, E., and Rapp, F. Immunologic Manipulation of Metas- tases Due to Herpes Virus Transformed Cells. Science, 180: 79-81, 1973.

19. Ennis, F. A. Host Defense Mechanisms against Herpes Simplex Virus. II. Protection Conferred by Sensitized Spleen Cells. J. Infect. Diseases, 127: 632-638, 1973.

20. Ennis, F. A., and Wells, M. Immune Control of Herpes Simplex Virus Infections. Cancer Res., 34: 1140-1145, 1974.

21. Fisher, G. W., Podgore, J. K., Balk, M. W., and Bass, J. W. Immunopo- tentiation and Antiviral Chemotherapy in Experimental Herpesvirus En- cephalitis. Pediat. Res. 9: 340, 1975.

22. Foley, F. D., Greenawald, K. A., Nash, G., and Pruitt, B. A. Herpesvirus Infection in Burned Patients. New Engl. J. Med., 282: 652-656, 1970.

23. Glasgow, L. A., and Habel, K. Role of Polyoma Virus and Interferon in a Herpes Simplex Infection in Vitro. Virology, 19: 328-339, 1963.

24. Glezen, W. P., Fernald, G. W., and Lohr, J. A. Acute Respiratory Disease of University Students with Special Reference to the Etiologic Role of Herpesvirus Hominis. Am. J. Epidemiol., 101: 111-121,1975.

25. Godfrey, R. C. Resistance to Intracerebral Challenge in Mice Immunized against Herpes Simplex Virus. Brit. J. Exptl. Pathol. 53: 529-538, 1972.

26. Gruia, M., Mutiu, C. A., Surdan, L., Gologan, R., Lupu, R., and Wecsler, P. The Action of Human Immunoglobulins in Experimental Herpes Infections in the Newborn Rat. Rev. Roumaine Virol., 10: 47-58, 1973.

27. Heineman, H. S. Herpes Simplex Neutralizing Antibody-quantitation of the Complement-dependent Fraction in Different Phases of Adult Hu- man Infection. J. Immunol., 99: 214-222, 1966.

28. Hirsch, M. S., Zisman, B.,and Allison, A. C. Macrophages and Age- dependent Resistance to Herpes Simplex Virus in Mice. J. Immunol., 104: 1160-1165, 1970.

29. Hollinshead, A. In: Conference and Workshop on Cellular Immune




Reactions to Human Tumor-Associated Antigens. Natl. Cancer Inst. Monograph, 37: 206, 1973.

30. Hooks, J. J., Fujibayashi, T., and Notkins, A. L. Production of Interferon by Immune Lymphocytes Exposed to Herpes Simplex Virus-Antibody Complexes. Federation Proc., 34: 869, 1975.

31. Johnson, R. T. The Pathogenesis of Herpes Virus Encephalitis. I1. A Cellular Basis for the Development of Resistance with Age. J. Exptl. Med., 120: 359-373, 1964.

32. Josey, W., Nahmias, A., and Naib, Z. Genital Herpes Simplex Infection: Present Knowledge and Possible Relationship to Cervical Cancer. Am. J. Obstet. Gynecol., 101: 718-729, 1968.

33. Kleinerman, E. S., Synderman, R., and Daniels, C. A. Depression of Human Monocyte Chemotaxis by Herpes Simplex and Influenza Vi- ruses, J. Immunol., 113: 1562-1567, 1974.

34. Kleinman, L. F., Kibrick, S. Ennis, F., and Polgar, P. Herpes Simplex Virus Replication in Human Lymphocyte Cultures Stimulated with Phy- tomitogens and Anti-lymphocyte Globulin. Proc. Soc. Exptl. Biol. Med., 141: 1095-1099, 1972.

35. Kretschmer, R., August, C. S., Rosen, F. S., and Janeway, C. A. Recur- rent Infections, Episodic Lymphopenia and Impaired Cellular Immunity. New Engl. J. Med., 281: 285-290, 1969.

36. Larson, C. L., Ushijima, R. N., Karim, R., Baker, M. B., and Baker, R. E. Herpesvirus Hominis Type 2 Infections in Rabbits: Effect of Prior Immu- nization wi th Attenuated Mycobacterium bovis (BCG) Cells. Infect. Im- munity, 6: 465-468, 1972.

37. Lausch, R. N., Jones, C., Christie, D., Hay, K., and Rapp, F. Spleen Cell- mediated Cytotoxicity of Hamster Cells Transformed by Herpes Simplex Virus: Evidence for Virus-specific Membrane Antigen. J. Immunol., 114: 459-465, 1975.

38. Lausch, R. N., Swyers, J. S., and Kaufman, H. E. Delayed Hypersensitiv- ity to Herpes Simplex Virus in the Guinea Pig. J. Immunol., 96:981-987, 1966.

39. Lehner, T., Shillitoe, E. J., Wilton, J. M. A., and Ivanyi, L. Cell-mediated Immunity to Herpes Virus Type 1 in Carcinoma and Pre-Cancerous Lesions. Brit. J. Cancer, 28 (Suppl. 1): 128-133, 1973.

40. Lerner, M. A., Lauter, C. B., Nolan, D. C., and Shippey, M. J. Passive Hemagglutinating Antibodies in Cerebrospinal Fluids in Herpesvirus Hominis Encephalitis. Proc. Soc. Exptl. Biol. Med., 140: 1460-1466, 1972.

41. Lerner, M. A., Shippey, M. J., and Crane, L. R. Serologic Responses to Herpes Simplex Virus in Rabbits: Complement-requiring Neutralizing, Conventional Neutralizing, and Passive Hemagglutinating Antibodies. J. Infect. Diseases, 129: 623-836, 1974.

42. Levi, M. M. Antigenicity of Ovarian and Cervical Malignancies with a View toward Possible Immunodiagnosis. Am. J. Obstet. Gynecol., 109: 889-698, 1971.

43. Linnemann, C. C., May, D. B., Schubert, W. K., Caraway, C. T., and Schiff, G. M. Fatal Viral Encephalitis in Children with X-linked Hypogam- maglobulinemia. Am. J. Diseases Children, 126: 100-103, 1973.

44. Lodmell, D. L., Niwa, A., Hayashi, K., and Notkins, A. L. Prevention of Cell-to-Cell Spread of Herpes Simplex Virus by Leukocytes. J. Exptl. Med., 137: 706-720, 1973.

45. Lodmell, D. L., and Notkins, A. L. Cellular Immunity to Herpes Simplex Virus Mediated by Interferon. J. Exptl. Med., 140: 764-778, 1974.

46. Logan, W. S., Tindall, J. P., and Elson, M. L. Chronic Cutaneous Herpes Simplex. Arch. Dermatol., 103: 606-614, 1971.

47. London, W. T., Catalano, L. W., Nahmias, A. J., Fuccillo, D. A., and Sever, J. L. Genital Herpesvirus Type 2 Infection of Monkeys. Obstet. Gynecol., 37: 501-509, 1971.

48. Lopez, C., and O'Reilly, R. Genetics of Natural Resistance of Mice to Herpes Virus Infection. Federation Proc., 34: 869, 1975.

49. Lycke, E., Hermadsson, S., Kristensson, K., and Roos, B. E. The Herpes Simplex Virus Encephalitis in Mice at Different Environmental Tempera- tures. Acta Pathol. Microbiol. Scand. B, 79: 502-510, 1971.

50. Merigan, T. C., and Stevens, D. A. Viral Infections in Man Associated with Acquired Immunological Deficiency States. Federation Proc., 30: 1858-1864, 1971.

51. Meyers, R. L., and Chitjian, P. A. Role of Immune Mechanisms in Control of Herpes Simplex Virus (HSV) Infections. Federation Proc., 34: 947, 1975.

52. Meyers, R. L., and Pettit, T. H. Corneal Immune Response to Herpes Simplex Virus Antigens. J. Immunol., 110: 1575-1590, 1973.

53. Meyers, R. L., and Pettit, J. H. The Pathogenesis of Corneal Inflamma- tion Due to Herpes Simplex Virus. I. Corneal Hypersensitivity to HSV Antigens in the Rabbit. J. Immunol. 111: 1031-1042, 1973.

54. Montgomerie, J. Z., Becroft, D. M. O., Croxson, M. C., Doak, P. B., and North, J. D. K. Herpes Simplex Virus Infection after Renal Transplanta- tion. Lancet, 2: 867-871, 1969.

55. Mori, R., Tasaki, T., Kimura, G., and Takeya, K. Depression of Acquired Resistance against Herpes Simplex Virus Infection in Neonatally Thy- mectomized Mice. Arch. Ges. Virusforsch., 2: 459-462, 1967.

56. Muller, S. A., Herrmann, E. C., and Winkelmann, R. K. Herpes Simplex

I m m u n o l o g y o f H S V I n f e c t i o n

Infections in Hematologic Malignancies. Am. J. Med., 52: 102-114, 1972.

57. Munoz, N. Effect of Herpesvirus Type 2 and Hormonal Imbalance on the Uterine Cervix of the Mouse. Cancer Res., 33: 1504-1508, 1973.

58. Nahmias, A., Fritz, M., and Chang, G. Herpesviruses as Infect ious and Oncogenic Agents in Man and Other Vertebrates. In: S. B. Day and R. A. Good (eds.), Membranes and Virus in Immunopathology, pp. 293-318. New York: Academic Press Inc., 1972.

59. Nahmias, A., Kibrick, S., and Rosan, R. C. Viral-leukocyte Interrelat ion- ships. I. Multiplication of a DNA Virus--Herpes S imp lex - - i n Human Leukocyte Cultures. J. Immunol., 93: 69-74, 1964.

60. Nahmias, A., and Roizman, B. Herpes Simplex Virus. New Engl. J. Med., 289: 667-674, 719-725, 781-789, 1973.

61. Nahmias, A. J., Alford, C., and Korones, S. Infection of the Newborn with Herpesvirus Hominis. Advan. Pediat., 17: 185-226, 1970.

62. Nahmias, A. J., Hirsch, M. J., Kramer, J. H., and Murphy, F. A. Effect of Antithymocyte Serum on Herpesvirus Hominis (Type 1 ) Infect ion in Adult Mice. Proc. Soc. Exptl. Biol. Med., 132: 696-698, 1989.

63. Nahmias, A. J., Josey, W. E., Naib, Z. M., Luce, C. F., and Guest, B. A. Antibodies to Herpesvirus Hominis Types 1 and 2 in Humans. I. Patients with Genital Herpetic Infections. Am. J. Epidemiol., 91: 547-552, 1970.

64. Nahmias, A. J., Naib, Z. M., Josey, W. E., and Clepper, A. C. Herpes Simplex Infection of the Female Genital Tract: Virological and Cytologi- cal Studies. Obstet. Gynecol., 29: 395-400, 1967.

65. Nahmias, A. J., Naib, Z. M. Josey, W. E., Franklin, E., and Jenkins, R. Prospective Studies of the Association of Genital Herpes Simplex Infec- tion and Cervical Anaplasia. Cancer Res., 33: 1491-1497, 1973.

66. Nahmias, A. J., Visintine, A., Reimer, C., Del Buono, I., Shore, S., and Starr, S. Herpes Simplex Virus Infection of the Fetus and Newborn. National Foundation Symposium on Fetus and Newborn, in press.

67. Nalick, R. H., DiSaia, P. J., and Rea, J. H. Immunological Response in Gynecological Malignancy as Demonstrated by the Delayed Hypersensi- tivity Reaction: Clinical Correlations. Am. J. Obstet. Gynecol., 118: 393- 405, 1974.

68. Notkins, A. L. Infectious Virus-antibody Complexes: Interact ion with Anti-lmmunoglobulins, Complement and Rheumatoid Factor. J. Exptl. Meal., 134: 419-519, 1971.

69. O'Reilly, R. I., and Lopez, C. Cyclic Deficiency of Lymphocyte Produc- tion in Patients with Recurrent Herpes Labialis or Progenital is. Federa- tion Proc., 34: 947, 1975.

70. Palmer, A., London, W., Nahmias, A., Naib, Z., Tunca, J., Fuccil lo, D., Ellenberg, J., and Sever, J. A Preliminary Report on Invest igat ion of the Oncogenic Potential of Herpes Simplex Virus Type 2 in Cebus Monkeys. Cancer Res., 36: 807-809, 1976.

71. Pien, F. D., Smith, T. F., Anderson, C. F., Webel, M. L., and Taswell, H. F. Herpesviruses in Renal Transplant Patients. Transplantat ion, 16: 489-495, 1973.

72. Price, R., Chernick, N. L., Horta-Barbosa, L., and Posner, J. B. Herpes Simplex Encephalitis in an Anergic Patient. Am. J. Med., 54: 222-228, 1973.

73. Rager-Zisman, B., and Bloom, B. Immunological Destruct ion of Herpes Simplex Virus 1 Infected Cells. Nature, 251: 541-542, 1974.

74. Rasmussen, L. E., Jordan, G. W., Stevens, D. A., and Merigan, T. C. Lymphocyte Interferon Production and Transformation after Herpes Simplex Infections in Humans. J. Immunol., 112: 728-736, 1964.

75. Rogers, H. W., Scott, V., and Patrode, R. A. Sensitization of Guinea Pigs to Herpes Simplex Virus. J. Immunol., 109: 801-806, 1972.

76. Rosenberg, G. L., Farber, P. A., and Notkins, A. L. In Vitro Stimulat ion of Sensitized Lymphocytes by Herpes Simplex Virus and Vaccinia Virus. Proc. Natl. Acad. Sci. U. S., 69: 756-762, 1972.

77. Rosenberg, G. L., and Notkins, A. L. Induction of Cellular Immunity to Herpes Simplex Virus: Relationship to the Humoral Immune Response. J. Immunol., 112: 1019-1025, 1974.

78. Rosenberg, G. L., Synderman, R., and Notkins, A. L. Product ion of Chemotactic Factor and Lymphotoxin by Human Leukocytes Stimulated with Herpes Simplex Virus. Infect. Immunity, 10: 111-115, 1974.

79. Rosenberg, G. L., Wohlenberg, C., Nahmias, A. J., and Notkins, A. L. Differentiation of Type 1 and Type 2 Herpes Simplex Virus by in Vitro Stimulation of Immune Lymphocytes. J. Immunol., 109: 413-414, 1972.

80. Rudneva, A., Korabelhikora, N. I., Germanov, A. B., Sokolcv, M. I., and Fadeeva, L. L. Studies of Interferogenic Activity of Herpes Simplex Virus. Arch. Virusforsch., 37: 1-5, 1972.

81. Russell, A. S. Cell-mediated Immunity to Herpes Simplex Virus in Man. Am. J. Clin. Pathol., 60: 825-829, 1973.

82. Russell, A. S. Cell-mediated Immunity to Herpes Simplex Virus in Man. J. Infect. Diseases, 129: 142-146, 1974.

83. Russell, A. S., Percy, J. S., and Kovithavongs, T. Cell-mediated Immu- nity to Herpes Simplex in Humans: Lymphocyte Cytotoxici ty Measured by ~lCr Release from Infected Cells. Infect. Immunity, 11: 355-359, 1975.

84. Russell, A. S., and Schlant, J. HL-A Transplantation Ant igens in Sub- jects Susceptible to Recrudescent Herpes Labialis, in press.

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A. J. N a h m i a s e t a l .

85. Shelley, W. B. Herpes Simplex Virus as a Cause of Erythema Multi- forme. J. Am. Med. Assoc., 201: 153-156, 1967.

86. Shore, S., Starr, S., Wood, P., and Nahmias, A. Detection of Cell- dependent Cytotoxic Antibody to Cells Infected with Herpes Simplex Virus, Nature, 251: 350-352, 1974.

87. Shore, S. L., Milgrom, H., Wood, P. A., and Nahmias, A. J. Efficacy of Cord Blood Mononuclear Cells in Antibody-dependent Cell Mediated Cytotoxicity to Target Cells Infected with Herpes Simplex Virus. Pediat. Res., 9: 335, 1975.

88. Simmons, R. L., Centifanto, Y., and Kaufman, H. E. Plaque-size Reduc- tion as a Measure of Viral Cell-mediated Immunity. Infect. Immunity, 10:1034-1039, 1974.

89. Spencer, E. S., and Anderson, H. K. Clinically Evident, Non-Terminal Infections with Herpesviruses and the Wart Virus in Immunosuppressed Renal AIIograft Recipients. Brit. Med. J., 3: 251-254, 1970.

90. Sprecher-Goldberger, S., Thiry, L., and Vandenbussche, P. Use of Immune Lymphocytes to Detect Expression of Herpetic Genome. Na- ture, 250: 678-679, 1974.

91. Start, S. E., Karatela, S. A. Shore, S. L., Duffey, A., and Nahmias, A. J. Stimulation of Human Lymphocytes by Herpes Simplex Virus Antigens. Infect. Immunity, 11: 109-112, 1975.

92. Stevens, J. G., and Cook, M. L. Restriction of Herpes Simplex Virus by Macrophages. An Analysis of the Cell-virus Interaction. J. Exptl. Med., 133: 19-38, 1971.

93. Stevens, J. G., and Cook, M. L. Maintenance of Latent Herpetic Infec- tion: An Apparent Role for Anti-viral IgG. J. Immunol., 113: 1685-1693, 1974.

94. St. Geme, J. W., Jr., Prince, J. T., Burke, B. A., and Good, R. A. Impaired Cellular Resistance to Herpes-Simplex Virus in Wiskott- Aldrich Syndrome. New Engl. J. Med., 273: 229-234, 1965.

95. StrOm, J. Herpes Simplex Virus as a Cause of Allergic Mucocutaneous Reactions (Ectodermosis Erosiva Pluriorificialis, Stevens-Johnson's Syndrome, etc.) and Generalized Infection. Scand. J. Infect. Diseases, 1: 3-10, 1969.

96. Sydiskis, R. J., and Schultz, I. Interferon and Antibody Production in Mice with Herpes Simplex Skin Infections. J. Infect. Diseases, 116: 455- 460, 1966.

97. Synderman, R., Wohlenberg, C., and Notkins, A. L. Inflammation and Viral Infection: Chemotactic Activity Resulting from the Interaction of Antiviral Antibody and Complement with Cells Infected with Herpes Simplex Virus. J. Infect. Diseases, 126: 207-209, 1972.

98. Templeton, A. Generalized Herpes Simplex in Malnourished Children. J. Clin. Pathol., 23, 24-30, 1970.

99. Thiry, L., Sprecher-Goldberger, S., Fassin, Y., Gould, I., Gompel, C., Pestiau, J., and Dehalleux, F. Variations of Cytotoxic Antibodies to Cells

with Herpes Simplex Virus Antigens in Women with Progressing or Regressing Cancerous Lesions of the Cervix. Am. J. Epidemiol., 100: 251-261, 1974.

100. Thong, Y. H., Vincent, M. M., Hensen, S. A., Fuccillo, D. A., Rola- Pleszczynski, M., and Bellanti, J. A. Depressed Specific Cell-mediated Immunity to Herpes Simplex Virus Type 1 in Patients with Recurrent Herpes Labialis. Infect. Immunity, 12: 76-80, 1975.

101. Tokamaru, T. A Possible Role of ~,A-Immunoglobulin in Herpes Simplex Virus Infection in Man. J. Immunol., 97: 248-259, 1966.

102. Valle, M. J., Bobrove, A. M., Strober, S., and Merigan, T. C. Immune Specific Production of Interferon by Human T Cells in Combined Macro- phage-lymphocyte Cultures in Response to Herpes Simplex Antigen. J. Immunol., 114: 435-441, 1975.

103. Valle, M. J., Jordan, G. W., Haahr, S., and Merigan, T. C. Characteris- tics of Immune Interferon Produced by Human Lymphocyte Cultures Compared to Other Human Interferons. J. Immunol., 115: 230-233, 1975.

104. Wallis, C., and Melnick, J. L. Herpesvirus Neutralization: The Rote of Complement. J. Immunol., 107: 1235-1242, 1971.

105. Walz, M. A., Price, R. W., Wohlenberg, C. R., and Notkins, A. L. Efficacy of Active Immunization upon the Development of a Latent Ganglion Infection with Herpes Simplex (HSV). Federation Proc., 34: 948, 1975.

106. Weintraub, I., Klisak, I., Lagasse, L. D., and Byfield, J. E. Evidence for Specific Tumor Cytotoxic Antibodies in Serum of Cervical Cancer Pa- tients. Am. J. Obstet. Gynecol., 116: 985-992, 1973.

107. Westmoreland, D., and Watkins, J. F. The IgG Receptor Induced by Herpes Simplex Virus: Studies Using Radioiodinated IgG. J. Gen. Virol., 24: 167-178, 1974.

108. Westmoreland, D., Watkins, J. F., and Rapp, F. Demonstration of a Receptor for IgG in Syrian Hamster Cells Transformed with Herpes Simplex Virus. J. Gen. Virol., 25: 167-170, 1974.

109. Wilton, J. M. A., Ivanyi, L., and Lehner, T. Cell-mediated Immunity in Herpesvirus Hominis Infections. Brit. Med. J., I : 723-726, 1972.

110. Yamamoto, Y. A Re-evaluation of the Skin Test of Herpes Simplex Virus. Japan J. Microbiol., 10: 67-77, 1966.

111. Yoshino, K., and Kiski, T. Studies on the Neutralization of Herpes Simplex Virus. VI. The Mode of Action of Complement upon Antibody Sensitized Virus. Japan J. Microbiol., 17: 63-70, 1973.

112. Yoshino, K., and Tanigushi, S. Evaluation of the Demonstration of Complement-requiring Neutralizing Antibody as a Means for Early Diag- nosis of Herpes Virus Infections. J. Immunol., 96: 196-203, 1966.

113. Zisman, B., Hirsch, M. S., and Allison, A. C. Selective Effects of Anti- macrophage Serum, Silica and Anti-lymphocyte Serum on Pathogen- esis of Herpes Virus Infection of Young Adult Mice. J. Immunol., 104: 1155-1159, 1970.




1976;36:836-844. Cancer Res André J. Nahmias, Steven L. Shore, Steve Kohl, et al. Herpes Simplex Virus Vaccines and Cervical CancerImmunology of Herpes Simplex Virus Infection: Relevance to

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Immunology of Herpes Simplex Virus Infection: …...[CANCER RESEARCH 36, 836-844, February 1976] Immunology of Herpes Simplex Virus Infection: Relevance to Herpes Simplex Virus Vaccines