Aging and Immunity II

Campus NovartisSiena - Italy

April 22-24, 2012

Steering Committee:

•Teresa AGUADO

• Giuseppe DEL GIUDICE

•Catherine DUTEL • Jörg GORONZY

• Beatrix GRUBECK-LOEBENSTEIN

• Agnes HOFFENBACH

• Jacques LOUIS

• Sandrine SAMSON

Prolongation of life expectancy in the 20th century has been one of the humanity’s greatest triumphs.The significant increase in human longevity however raises the crucial issue of healthy aging. Since infectious diseases significantly contribute to morbidity in the particularly vulnerable elderly population, strategies for preventing these diseases would have a clear impact on improving healthy aging. However, given the age-induced variations of the immune responsiveness, vaccines and immunization strategies tailored for the elderly population are needed and should be developed.

These issues were reviewed and discussed during a meeting organized by Fondation Mérieux in Siena in September 2009. During this meeting, the several possible causes of immune senescence were reviewed, and the strategies for counteracting this waning of immune responsiveness and for restoring immunocompetence were discussed. In addition, examples of diseases that should be targeted by vaccination in the senior population were considered.

Thus, inasmuch as aging represents one of the main health problems of the 21st century, vaccines specifically directed to the elderly become a public health priority both in developed and in developing countries. Within this context, Fondation Mérieux is organizing this second meeting on ‘Aging and Immunity’ with the objectives to review the progress made during the last 2 and ½ years. During this meeting, particular attention will be given to discussing the advancement made in the high priority issues identified by participants to the 2009 workshop.

These issues include:

- Assessing the progress in the understanding of the cellular and molecular basis for immune senescence. Strategies for strengthening the immune reactivity of the elderly to vaccination without triggering potentially adverse inflammatory reactions will be considered.- Reviewing the evidence that vaccination in the elderly delays the transition from « young old » to « old old », and identifying the age groups within the elderly population that should be particularly targeted by vaccination.- Discussing the needs for reliable and robust biomarkers of vaccine-induced protection in the elderly (correlates of protective immunity) and for better diagnostic tools.- Defining strategies by which progress made in understanding the basis of immunosenescence can be integrated with vaccinology and gerontology, and the most appropriate ways to translate the current scientific knowledge into the development of vaccination strategies for the poorest countries.

Background

ScientificProgramme

Sunday 22 April 2012

15.00 - 16.00 Registration and welcome coffee

16.00 - 16.30 Welcome Address

Rino RAppuolI(Global Head, Vaccines Research, Novartis Vaccines & Diagnostics) & Fondation Mérieux

16.30 - 17.10Keynote lecture:Strategies for making vaccines efficaciousintheelderly

Rino RAppuolI

Session 1 The burden of infectious diseases in the elderly17.10 - 18.30 Chair: Giuseppe Del GIuDICe

17.10 - 17.30 Burden and relative importance of infectious diseases in the elderly Karl-Heinz KRAuSe

17.30 - 17.50 Discussion

17.50 - 18.10 WHO initiatives in relation to aging and immunity Teresa AGuADo

18.10 - 18.30 Discussion

ScientificProgramme

08.30 - 08.50Response of the aged immune system to acute and persistent infection

Janko NIKolICH-ZuGICH

08.50 - 09.10 Discussion

09.10 - 09.30 Signaling pathways in the aged CD4 T cell Jörg GoRoNZy

09.30 - 09.50 Discussion

09.50 - 10.20 Coffee break

10.20 - 10.40 CD8+ T cell response in the elderly Beatrix GRubeCK-loebeNSTeIN

10.40 - 11.00 Discussion

11.00 - 11.20 B cell repertoire and aging Deborah DuNN-WAlTeRS

11.20 - 11.40 Discussion

11.40 - 12.00 Age associated alterations in human dendritic cell function Albert SHAW

12.00 - 12.20 Discussion

12.30 - 14.00 Lunch

14.00 - 14.20 The alteration of Cutaneous T Cell immunosurveillance during ageing Arne AKbAR

14.20 - 14.40 Discussion

14.40 - 15.00 HIVinfection,inflammation,immunosenescence and aging Steven G. DeeKS

15.00 - 15.20 Discussion

Monday 23 April 2012

08.15 - 08.30 Welcome coffee

Session2 Progressintheunderstandingofthemechanismsunderlying immunosenescence08.30 - 15.20 Chair: beatrix GRubeCK-loebeNSTeIN

ScientificProgramme

Session3 Newtechnologiestoredefinebiomarkersofimmuneresponsiveness in the elderly15.20 - 17.50 Chair: Jörg GoRoNZy

15.20 - 15.40 Many new dimensions in single cell proteomics Wendy FANTl

15.40- 16.00 Discussion

16.00 - 16.30 Coffee break

16.30 - 16.50

A system biology approach to predict immunogenicity and provide new mechanistic insights about vaccines

Bali PuleNDRAN

16.50 - 17.10 Discussion

17.10 - 17.30Computational biology to evaluate genes expression following vaccination

Damien CHAuSSAbel

17.30 - 17.50 Discussion

Tuesday 24 April 2012

Session 3 Continued08.30 - 09.50

08.30 - 08.50 Deep sequencing of B cell repertoire and aging Scott boyD

08.50 - 09.10 Discussion

09.10 - 09.30Dissecting the human T cell response to pathogens and vaccines

Federica SAlluSTo

09.30 - 09.50 Discussion

ScientificProgramme

09.50 - 10.10 The needs of the aging population with new vaccines Martine DeNIS

10.10 - 10.30 Discussion

10.30 - 11.00 Coffee break

11.00 - 11.20Age-related differences in the antibody responses to seasonal and pandemicinfluenzavaccines

Harry GReeNBeRG

11.20 - 11.40 Discussion

11.40 - 12.00 Adjuvants for the elderly Giuseppe Del GIuDICe

12.00 - 12.20 Discussion

12.20 - 12.40Pneumoccocalvaccinesinthe elderly (conjugated vs polysaccharides)

Paolo boNANNI

12.40 - 13.00 Discussion

13.00 - 14.30 Lunch

14.30 - 14.50 Herpeszostervaccinesfortheelderly Myron leVIN

14.50 - 15.10 Discussion

Session 5 Round table discussion and closing of the meeting15.10 - 16.30

15.10 - 16.30 Discussion on future development and needs

Session4 Improvingvaccineefficacyintheelderly09.50 - 15.10 Chair: Teresa AGuADo

Session 1

The burden of infectious diseases in the elderly

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Burden and relative importance of infectious diseases in the elderly

Karl-Heinz KRAuSeGeneva university Hospitals - Switzerland

Although demographic aging does not remain restricted to industrialized countries, the medical challenge arising from the aging population will be distinct in the developing world. This is particularly true with respect to infectious diseases, which have a distinct spectrum in the elderly population, as well as a greater overall relevance in the developing world. Tropical diseases have a specific presentation and epidemiology in elderly patients. Infectious diseases with a worldwide distribution impact elderly patients in the developing world in a specific manner, which is most obvious with respect to human immunodeficiency virus and tuberculosis but is also true with respect to «trivial» manifestations of infection, such as diarrhea and pneumonia. Malnutrition contributes in a major way to the immunodeficiency of elderly patients in the developing world. Poorly controlled use of antimicrobial drugs leads to multidrug-resistant microorganisms, which, together with the limited resources available for drug treatment, makes appropriate treatment of infections in elderly patients in developing countries very difficult. Infections in elderly patients will have an increasing impact on the public health and economy of developing countries.

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WHO initiatives in relation to ageing and immunity

Teresa AGuADoJudith Thomas-CrusellsWorld Health organization - Switzerland

The improvement of child survival, reduced mortality rates and decreased fertility rates worldwide are all factors contributing to the wellbeing of mankind. However, combined with increased longevity, one consequence is the ageing of the global population. By the year 2030 the percentage of the population that will be elderly (≥ 60y of age) is predicted to represent over 25% of the total population, of whom 75% will be living in less developed countries.

Vaccines are one of the most cost-effective health interventions, and have proven to be crucial in preventing mortality and morbidity linked to a large number of pathogens. While vaccination of infants remains an unfinished task, sustained protection against infectious diseases in adulthood and older ages should become a goal of a comprehensive health program.

Therefore a life-course approach to immunization as a novel goal deserves insight and the potential impact on quality of life in older age should be fully explored. If one is to anticipate the future needs in this regard, our efforts should be global, as population studies indicate a major expansion of aged populations everywhere in the world. Information exists on aged populations and infection, as well as data on impaired immune responses as life progresses, but the information is fragmented and mostly collected in the high income countries of the world. The middle and low income settings remain largely undocumented.

The World Health organization (WHo), through a joined effort of their ageing and life-course (AlC) and immunization departments (IVb/IVR), aims to develop a research agenda on the use of vaccines and immunization during the life course of an individual, which could eventually result in increased quality of life and significant savings to the health system.

Developing such a research agenda requires combining findings on populations and gerontology with immunology and molecular biology, in order to generate knowledge that eventually will lead to policy for the vaccination of groups beyond infancy. Data should be collected in populations of different ages and of different areas of the world, so the information generated can lead to global policy.

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Continued

In the past years several meetings either convened by WHo or jointly organized with other groups helped us to start understanding the existing information and the needs of the field and in turn, to provide some guidance for a broad research agenda. As a first step, and based on the recommendations of an ad-hoc group of experts, WHo has identified some main work streams/areas of work to devote efforts to studies in the following areas:

• epidemiology and disease burden • Immunological status/immune senescence and related methodology• Implementation/programmatic research with selected vaccines• Socio-economic analysis to assess and compare benefits of different interventions

A number of activities have been prioritized and some are already ongoing, related to the first two bullet points: 1) A global systematic review of disease burden (eg respiratory infections), with emphasis on morbidity throughout the life of an individual has been initiated. 2) Methodology to assess the immunological status (eg T cell status) of a major collection of dried blood samples (several thousand samples) from individuals from all regions of the world and of different ages is being validated. The immediate step will be to apply the measurement to a representative part of the collection to assess the effective resting naïve T-cell pool (marker of ageing).3) A network of experts should be established in order to coordinate their studies for an optimal understanding of the process of immune senescence.

Session 2

Progressintheunderstandingofthe mechanisms underlying immunosenescence

Session 2

Progressintheunderstandingofthe mechanisms underlying immunosenescence

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Response of the aged immune system to acute and persistent infection in mice and humans

Janko NIKolICH-ZuGICHDepartment of Immunobiology and the Arizona Center on Aging university of Arizona College of Medicine - uSA

Defense against intracellular infections is chiefly mediated by T-cells, a cell type severely affected by the process of aging. We have studied T-cell subset-specific responses in aging and have placed them in the context of acute or latent persistent infection to elucidate how immune defense to such infections may change with aging. We have discovered numerous defects that cumulatively affect: (i) the ability to detect, process and present antigens to T cells; (ii) the ability of naïve old T cells to be activated, proliferate and differentiate into robust effector cells; (iii) the ability of homeostatic mechanisms to maintain and preserve T-cell subset balance; and (iv) the ability to do all of the above while dealing with a latent persistent herpesvirus infection. I will discuss essential parallels between the findings in mice and humans in all of the above defects and will conclude by reviewing our efforts to rationally correct each of the problems, with the goal to eradicate infectious diseases as a cause of death in aging.

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Signaling pathways in the aged CD4 T cell

Jörg J. GoRoNZyStanford university School of Medicine - uSA

Alterations in signaling pathways seen in aging CD4 T cells frequently build on mechanisms that are operational in T cell development and T cell differentiation. Calibration of T cell receptor (TCR) signaling thresholds is an essential element in the life cycle of a T cell. Most notably, thymocytes have an exquisite TCR sensitivity which allows them to be positively selected on self-antigens. The naive T cell that leaves the thymus is no longer able to respond to self-antigens with a productive response. The altered signaling threshold is caused by a decline in miR181a that controls the expression of several phosphatases including the dual-specific phosphatase (DuSp) 5 and DuSp6. miR181a levels in naive and memory T cells continue to decline with age. Reconstitution of miR181a levels, or transcriptional silencing or pharmacological inhibition of DuSp6 markedly improves responsiveness of elderly T cells. other age-associated alterations in signaling feedback mechanisms represent adaptations to the changing environment in the aging cell. upon activation, aged CD4 memory T cells exhibit increased levels of DuSp4 that curtails the nuclear activity of phosphorylated eRK and JNK. The increased transcription of DuSp4 is due to lower energy production and increased AMpK activity. Silencing of DuSp4 expression in elderly CD4 T cells restores their ability to provide helper activity for B cell differentiation and antibody production. The identification of activated negative feedback loops involving several members of the DuSp family in aged CD4 T cells open the opportunity that signaling pathways in T cells can be directly targeted to optimize vaccine responses in the elderly.

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CD8+ T cell response in the elderly

Beatrix GRubeCK-loebeNSTeINInstitute for biomedical Aging Research of the Austrian Academy of Sciences - Austria

In the past we and others have demonstrated that aging leads to a shift in the CD8+ T cell repertoire from naïve to highly differentiated effector T cells. This shift is accelerated by latent infection with cytomegalovirus and thymectomy. We have now concentrated on the still controversial issue whether accumulation of effector CD8+ T cells is due to continuous regeneration or decreased degradation. We demonstrate that effector CD8+ T cells are specifically enriched in niches such as the bone marrow, where they reside in a state of pre-activation and can produce cytokines upon stimulation. This is not due to an intrinsic resistance to apoptosis-inducing stimuli as suggested by others, as we can also show that highly differentiated effector CD8+ T cells are particularly susceptible to apoptosis mediated by extrinsic and intrinsic factors. This susceptibility to apoptosis is at least partly due to a decreased capacity of the specific cell type to repair DNA damage. The proneness of effector CD8+ T cells to undergo apoptosis can be overcome by intensive Il-15 signaling, a characteristic feature of the aged bone marrow. Terminally differentiated effector CD8+ T cells may therefore represent a useful line of defense in old age in certain organs. This demonstrates that senescent cells can still fulfill important functions and compensate for the loss of regenerative capacity.

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B cell repertoire and ageing

Deborah DuNN-WAlTeRSKing’s College london School of Medicine - uK

We have previously shown that the b cell repertoire in the old is reduced, and hypothesised that this may contribute to the impaired humoral responses of older people. We have now investigated the repertoire and antibody responses to winter vaccination in two age groups, aged 18-49 and 65-89. We found that the serum IgM and IgA pneumococcal responses were significantly impaired in the older group, with no difference in IgG levels. Repertoire analysis by CDR3 spectratyping showed a clear change in the repertoire at day 7 after vaccination, with a return to the baseline levels at day 28. The changes at day 7 reflected expansion of IGH sequences that have smaller, more hydrophilic, CDR3 regions and these changes were attenuated in the older group. The older group were more likely to have spectratypes indicative of a reduced diversity at day 0 and day 28. on average the baseline repertoire in the older group was comprised of larger CDR3 regions than in the younger group. We have also seen age-related changes in IGHV gene usage, which may reflect changes in the selection and expansion events that take place during b cell development and the humoral immune response. our data shows that both activation and resolution of the b cell response can be affected by age, CDR3 regions are particularly important in the humoral immune response and the repertoires of some B cell sub-populations are more affected by age than others.

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Age-associated alterations in human dendritic cell function

Albert SHAWyale university - School of Medicine - uSA

Age-associated changes in immune function, termed immunosenescence, contribute to impaired vaccine responses and increased morbidity and mortality from infectious diseases in older adults. Studies of immunosenescence have focused on the adaptive immune system and alterations in B and T cell function. However, recent findings have indicated that immunosenescence occurs in the innate immune system as well. In particular, aging is associated with altered function of the Toll-like Receptor (TlR) family of innate immune pattern recognition receptors. evidence from our laboratory and others indicates that TlR-induced cytokine production in myeloid (mDC) and plasmacytoid (pDC) dendritic cell populations is decreased in cells from older (≥ 65 years of age), compared to young (age 21-30) adults. However, basal cytokine production in DCs from older, but not young individuals appears markedly elevated; this dysregulated cytokine production may not be able to be substantially augmented with additional TlR stimulation, while contributing to the paradoxical pro-inflammatory milieu associated with human aging. In monocyte-derived DCs, evidence for both increased and decreased cytokine production in response to TlR engagement or viral infection has emerged, suggesting further complexity in the consequences of aging. Recent studies from our laboratory also indicate that TlR-induced costimulatory protein expression is altered in DCs from older, compared to young adults. both TlR-induced cytokine production and costimulatory protein expression are strongly associated with antibody response to influenza vaccine, reinforcing the link between the innate and adaptive immune systems and highlighting the possibility of enhancing innate immune responses as a means to improve vaccine efficacy in older adults.

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The alteration of cutaneous T Cell immunosurveillance during ageing

Arne AKbARDivision of Infection and Immunity - university College london - uK

Immunity declines during aging, however the mechanisms involved in this decline are not known. In this study, we show that cutaneous delayed type hypersensitivity (DTH) responses to recall antigens are significantly decreased in older individuals. However, this is not related to CC chemokine receptor 4, cutaneous lymphocyte-asso-ciated antigen, or CD11a expression by CD4+ T cells or their phy-sical capacity for migration. Instead, there is defective activation of dermal blood vessels in older subject that results from decreased TNF-alpha secretion by macrophages. This prevents memory T cell entry into the skin after antigen challenge. However, isolated cutaneous macrophages from these subjects can be induced to secrete TNF-alpha after stimulation with Toll-like receptor (TlR) 1/2 or TlR 4 ligands in vitro, indicating that the defect is reversible. The decreased conditioning of tissue microenvironments by macro-phage-derived cytokines may therefore lead to defective immuno-surveillance by memory T cells. This may be a predisposing factor for the development of malignancy and infection in the skin during aging. Furthermore we demonstrate that there is a significant in-crease in the proportion of regulatory CD4+Foxp3+ T cells in the skin of older humans that may contribute the inhibition of macro-phage activation and TNF-alpha during ageing.

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HIV infection, inflammation, immunosenescence andaging

Steven G. DeeKSuniversity of California - uSA

Most HIV infected patients who are motivated to take therapy are now able to achieve durable and perhaps life-long viral suppression. As a consequence, the classic AIDS-related conditions are becoming less common. This does not mean that treated individuals have completely restored health. Indeed, there is a growing consensus that when compared to HIV uninfected persons, patients on highly active antiretroviral therapy (HAART) remain at increased risk for “non-AIDS” morbidity. Many of the complications are similar to those that occur during the normal aging process (e. g., cardiovascular disease, cancer). These complications are predicted by and possibly caused (in part) by HIV-associated immunologic dysfunction that is not reversed by HAART. This includes measures of immunologic aging, including chornic inflammation (variably defined), the frequency of memory effector CD8+CD28- T cells, the CD4/CD8 ratio, and T cell proliferation, all of which often remain abnormal during treated disease.

our group has performed extensive immunologic studies in large numbers of untreated and treated HIV infected adults. With regard to broadly defined characteristic of immunologic aging and immunosenescence, we have found that (1) T cell activation-as defined by co-expression of CD38 and HlA-DR—increase dramatically during acute HIV infection, long before onset of obvious immunodeficiency, (2) an “immune activation” set-point is established early in HIV disease that is associated with rate of subsequent CD4+ T cell declines, independent of the level of viremia, (3) immune activation decreases during long-term antiretroviral therapy but rarely returns to a normal level, (4) combination antiretroviral therapy often fails to reconstitute a normal CD4+ T cell count, even after up to 10 years of effective viral suppression, (5) CMV-specific T cell responses are at least twice a high in long-term treated HIV infection as compared to that observed in age matched HIV uninfected persons, (6) the level of CMV-specific T cells responses is strongly associated with markers of heart disease, (7) valganciclovir “intensification” in antiretroviral-treated disease reduces a substantial proportion of the excess levels of immune activation seen in these individuals, (8) compared to HIV-uninfected controls, antiretroviral-treated patients have lower frequency of naïve CD8+ T cells and a higher frequency of CD28- CD8+ T cells, and (9) the frequency of immunosenescent cells predicts degree of atherosclerosis in HIV infected women.

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Continued

There are number of questions which remain to addressed regarding HIV infection and immunosenescence, including: (1) does immunosenescence (as measured in blood and mucosal tissues) persist indefinitely during treated HIV infection (and if so, is this apparent in those who are middle-aged)?, (2) is there an easy way to define subset of patients who are at risk for failure to restore a normal immune system?, (3) what role if any does residual low level HIV replication and/or microbial translocation have on immunosenescence and vaccine responsiveness during antiretroviral therapy?, and (4) what is the in vivo significance of immunosenescence? Knowledge gained regarding these issues could provide needed rationale (and necessary biomarkers) for novel targeted interventions to reverse immunologic aging in this well-characterized group of patients. This knowledge could provide novel insights in the role of chronic inflammation in aging, and hence may have significance to more general questions pertaining to aging.

Session 3

Newtechnologiestoredefine biomarkers of immune responsiveness in the elderly

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Many new dimensions in single cell proteomics

Wendy FANTlSean C. bendall, erin F. Simonds, Kara l. Davis D.o, evan W. Newell, Natalia Sigal, Mark M. Davis & Garry p. Nolan

baxter laboratory for Stem Cell biology - Stanford university - uSA

elemental mass spectrometry-based cytometry (mass cytometry) is a recently developed technology in which fluorochromes have been replaced with elemental metal isotopes. These isotopes are attached to cells via cell affinity binders, labeled with polymeric, metal- loaded chelators. The quantity and mass of these stable isotopes are detected by means of an inductively-coupled plasma mass spectrometer (ICp-MS), which in the current generation of instruments allows for the analysis of spectra encompassing 100 isotope channels at an acquisition rate of 1000 cells/second. The high resolution of the time-of-flight (ToF) mass analyzer used in the mass cytometer combined with the intrinsically discrete nature of isotopic masses, allows discrimination of isotopes separated by only one atomic mass unit with negligible spectral overlap. This dramatically increases the number of parameters that can be measured simultaneously per single cell, especially when compared with fluorochrome-tagged reagents. Cell affinity binders such as antibodies, chemical linkers and most recently, tetramers have all been used successfully with this new technology.

Recently, we used mass cytometry to measure intracellular signaling responses mediated by extracellular modulators within immunophenotypic cell subsets. Cells from healthy individuals, spanning the hematopoietic continuum in bone marrow as well as peripheral blood mononuclear cells were investigated for 34 parameters. (bendall et al. Science 2011). The single cell data from this investigation represent the reference against which studies of drug action and disease states can be compared.

Two applications of mass cytometry will be presented in which an underappreciated level of cellular complexity is revealed. In the first, the human CD8+ cytotoxic T cell compartment was surveyed with specific peptide–MHC tetramers labeled with heavy metals, allowing the delineation of ebV, CMV and influenza specific CD8+ T-cell subsets.

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Continued

Simultaneously a panel of 26 antibodies against surface markers, cellular cytokines and granule components further characterized their specific functional attributes (Newell et al. Immunity 2012). In the second study, a 35 parameter single-cell analysis of diagnostic pediatric b-cell All samples revealed disease architecture based on immunophenotype while simultaneously identifying signaling networks within specific cell subsets. With subsequent drug and cytokine treatment of these cells, further information about their internal network organization was evident. The latter would not be revealed in the basal cell signaling state or with immunophenotypic analysis alone.

The greatly enhanced number of parameters measured by mass cytometry at the single cell level will afford new opportunities for improving patient outcomes to therapeutic intervention.

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Systems Vaccinology: enabling rational vaccine design with systems biology

bali puleNDRANuniversity of emory - uSA

Despite their great success, we understand little about how effective vaccines stimulate protective immune responses. Two recent developments promise to yield such understanding: the appreciation of the crucial role of the innate immune system in sensing microorganisms and tuning immune responses, and advances in systems biology. In this presentation, I will discuss how these developments are yielding insights into the mechanism of some of the most successful vaccines ever developed. Furthermore, such developments promise to address a major challenge\ in vaccinology: that the efficacy of a vaccine can only be ascertained retrospectively, upon infection. The identification of molecular signatures induced rapidly after vaccination, which correlate with and predict the later development of protective immune responses, would represent a strategy to prospectively determine vaccine efficacy. Such a strategy would be particularly useful when evaluating the efficacy or immunogenicity of untested vaccines, or inidentifying individuals with sub-optimal responses amongst high risk populations, such as infants or the elderly. We have recently used a systems biology approach to identify early gene signatures that correlate with, and predict the later immune responses in humans vaccinated with the live attenuated yellow fever vaccine yF-17D, or with the influenza vaccines. I will review these studies, and discuss their broader implications for vaccinology.

References

1. Systems vaccinology. pulendran b, li S, Nakaya HI. Immunity. 2010 oct 29;33(4):516-29.

2. Systems biology approach predicts immunogenicity of the yellow fever vaccine in humans. Querec TD, Akondy RS, lee eK, Cao W, Nakaya HI, Teuwen D, pirani A, Gernert K, Deng J, Marzolf b, Kennedy K, Wu H, bennouna S, oluoch H, Miller J, Vencio RZ, Mulligan M, Aderem A, Ahmed R, pulendran b. Nat Immunol. 2009 Jan;10(1):116-25. epub 2008 Nov 23.

3. Immunological mechanisms of vaccination. pulendran b, Ahmed R. Nat Immunol. 2011 Jun;12(6):509-17.

4. Systems biology of vaccination for seasonal influenza in humans. Nakaya HI, Wrammert J, lee eK, Racioppi l, Marie-Kunze S, Haining WN, Means AR, Kasturi Sp, Khan N, li GM, McCausland M, Kanchan V, Kokko Ke, li S, elbein R, Mehta AK, Aderem A, Subbarao K, Ahmed R, pulendran b. Nat Immunol. 2011 Jul 10;12(8):786-95. doi: 10.1038/ni.2067

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Computational biology to evaluate genes expression following vaccination

Damien CHAuSSAbelbenaroya Research Institute - uSA

The study of a system requires a detailed understanding of its constitutive elements but also a holistic understanding of how those elements interact to produce the complex molecular and cellular chain of events that give rise to immune responses. Widely available large-scale profiling platforms can capture the state of entire systems. But exploiting this information to gain in depth understanding of the organization of those systems remains a challenge. We have developed sophisticated computational approaches in order to simplify the analysis and interpretation of large-scale datasets. The resulting modular analysis frameworks can be used to develop novel tools for the assessment of the immune system in large-scale human immunology studies. This talk will illustrate the application of such methodologies for the study of disease pathogenesis and responses to vaccines.

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Deep sequencing of B cell repertoire and aging

Scott D. BoyDDepartment of pathology - Stanford university - uSA

Some elderly people have a compromised immune system with decreased functional capacity. To what degree this is associated with an altered immunoglobulin repertoire and changes in b cell functional responses is unclear. using high-throughput DNA sequencing, we have sequenced immunoglobulin heavy chain V(D)J rearrangements from healthy participants 1 to 88 years old. Some participants are sampled twice at two time points of one year apart. Sequence analysis shows little age-related change in V, D, J segment usage, V-D and D-J junction properties, or complementary determining region 3 (CDR3) features during adult life. Hypermutation frequency of the V genes in the total repertoire is also comparable between adult subjects of various ages. Clonal expansion analysis, however, reveals that limited numbers of large expanded clones are more frequently observed in elderly people than in younger adults. In addition, the large clones in elderly people distinguish themselves from those in younger people in that they appear to persist over time. Correlation of these results with chronic viral infection status, and the clinical entity b cell monoclonal lymphocytosis, is underway.

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Dissecting the human T cell response to pathogens and vaccines

Federica SAlluSToInstitute for Research in biomedicine - Switzerland

We have developed a high throughput cellular screening approach to interrogate the human naïve and memory T cell repertoires. The method is based on the polyclonal stimulation and expansion of limiting number of T cells in multiple replicate cultures to generate libraries that can be repeatedly screened for the presence of antigen specific T cells using antigen and autologous ApCs. I will present data obtained in the laboratory on the distribution in different memory subsets of T cells specific for microbial antigens, vaccines, allergens, and self-antigens.

Session 4

Improvingvaccineefficacyin the elderly

Session 4

Improvingvaccineefficacyin the elderly

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The needs of the aging population with new vaccines

Martine DeNISSanofi pasteur - France

Vaccination is the most straightforward approach to prevent infectious diseases and their sequelae in all age groups. Despite the availability of various vaccines, infectious diseases cause significant morbidity and mortality in elderly subjects. The situation could be improved by targeting the objectives below.

1°/ Develop new vaccines targeting diseases affecting elderly subjects.Clostridium difficile is the most frequent cause of nosocomial diarrhea. While hospital-acquired infections cause a considerable problem in most industrialized countries, C.difficile infections (CDI) are also increasingly reported in the community. elderly persons are at an increased risk of CDI-related morbidity and mortality. Treatment options are available but suboptimal. An association between increased serum levels of IgG antibody against toxin A and asymptomatic carriage of C. difficile provides a rationale for vaccine development. Sanofi pasteur’s candidate (currently in phase II of its clinical development) is a toxoid vaccine designed to neutralize the harmful effects of the toxins A and b, hence for the prevention of CDI.

2°/ Adapt existing vaccines to the elderly.Some vaccines have suboptimal efficacy when used in an elderly population. The reduced response to vaccination may result from the age-related impairment of immune functions and the increase in comorbidities. Various attempts have been made to improve the efficacy of influenza vaccines in elderly subjects. Changes in the vaccination schedule, vaccine dose, route of immunization as well as combination with an adjuvant or administration of a biological response modifier have been attempted with unequal success. Nevertheless, the available data demonstrate for selected approaches the feasibility of counterbalancing the effects of immunosenescence on the response to vaccination.

3°/ Increase vaccination coverage. Various surveys indicate that the coverage remains low for most recommended vaccines in adults. For instance, a recent study in Turkish farmers over 50 years of age reported detection of a seroprotective tetanus anti-toxin level in only 34% of study participants. better vaccine coverage will be needed to substantially reduce the occurrence of vaccine-preventable diseases in the elderly.

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Age-related differences in the antibody responses to seasonalandpandemicinfluenzavaccines

Harry b. GReeNBeRGStanford university - uSA

The limited efficacy of influenza vaccines in the elderly has been linked to reduced serum antibody responses. To investigate the mechanism for the differential antibody responses between elderly and young adults, we studied influenza vaccine-activated plasmablasts in the peripheral blood 7 days after immunization with either the inactivated 2009 seasonal TIV influenza vaccine or the 2009 monovalent pandemic H1N1 vaccine. We measured the numbers of vaccine-specific plasmablasts as well as the quantity of antibodies secreted by these cells. We also examined the specificity and affinity/avidity of the secreted antibodies against variant influenza strains. our data revealed distinct quantitative and qualitative characteristics of the plasmablast responsiveness in young versus elderly subjects as well as between the two different vaccine preparations. These results suggests that the diminished antibody response in elderly subjects is affected by a reduced capacity of b cell amplification, and also substantially by the memory B cell repertoire that is shaped by the age-associated influenza exposure history.

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Adjuvants for the elderly

Giuseppe Del GIuDICeGlobal Head Translational Medicine, Novartis Vaccines and Diagnostics - Italy

With the ageing of the population worldwide, the need for improved vaccines for the elderly is increasing significantly. Several approaches are being undertaken to counteract the reduced ability of elder population to respond properly to vaccination, with the final goal of offering better protection against infectious diseases. Among these approaches, particular attention has been received by adjuvants, and in particular by oil-in-water adjuvants, such as MF59, which have probably given so far the best results in improving immune responsiveness in elder individuals, both against the seasonal and against the pandemic influenza viruses, as well as against other microbial agents. This adjuvant not only enhances the immune response at T and b cell levels quantitatively, but it also dramatically affects the quality of the antibody response induced by the adjuvanted vaccines, as well as the repertoire of the b cell epitopes recognized by these antibodies. Much knowledge has been recently acquired on the early immunological events triggered by MF59. More work is in progress now to decipher the mechanisms by which it exerts its effects on both T and b cells at older ages. This knowledge, together with a deeper understanding of the mechanisms leading to the poorer responses in the elderly, would further help to better tune the development of stronger vaccines in these immunologically frail individuals.

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Pneumoccocalvaccinesintheelderly(conjugatedvspolysaccharides)

Paolo BoNANNIuniversity of Florence - Department of public Health - Italy

Pneumococcal polysaccharide vaccines have been used since a long time for the prevention of pneumococcal diseases in the elderly and in risk groups. Their efficacy and effectiveness has always been controversial. There is substantial agreement that the presently used 23-valent polysaccharide vaccine is 50-80% effective for the prevention of invasive pneumococcal diseases in healthy adult subjects. However, no definitive proof is available about its efficacy against pneumonia, although several data on the impact on hospitalizations due to pneumonia suggest a possible relevant effect.Recently, the 13-valent conjugate pneumococcal vaccine has been licensed by the eMA also for use in subjects aged ≥50 years, with indication against invasive pneumococcal diseases, based on comparative immunogenicity data with the polysaccharide vaccine. Immunogenicity was shown to be always non-inferior, and in most instances superior to that of the polysaccharide vaccine for the common serotypes.effectiveness data for the conjugate vaccine are not yet available, but are awaited within 1 year and a half from a big study of effectiveness on community acquired pneumonia presently underway in the Netherlands.public health decisions will have to be made in the near future about the best strategy of use of the two vaccines, that will have to take into account the current open questions on immunological, effectiveness, ethical, organizational and communication issues.

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Herpeszostervaccinesfortheelderly

Myron leVINuniversity of Colorado School of Medicine and Health Sciences Center - uSA

Varicella-zoster virus (VZV) T-cell-mediated immunity (VZV-CMI) in older persons prevents latent VZV in sensory neurons from reactivating and causing herpes zoster. VZV-CMI declines greatly with ageing. Specifically, there is a significant loss of VZV-specific CD4+ early effectors and CD4+ effector memory cells, and lower CD8+ early effectors. These can be restored by the licensed (and recommended) zoster vaccine. However, the vaccine-induced boost in VZV-CMI (and thus the efficacy of the vaccine) is a function of the age of the vaccinee, and the duration of the boost wanes with time. both factors influence the value of zoster vaccine. based on information from these and other studies some potential approaches to improving prevention of herpes zoster are discussed.

Teresa Aguado aguadom@who.int WHo Arne Akbar a.akbar@ucl.ac.uk university College london

Paolo Bonanni paolo.bonanni@unifi.it university of Florence, Department of public Health

Scott Boyd sboyd1@stanford.edu university of Stanford, Department of Pathology

Damien Chaussabel DChaussabel@benaroyaresearch.org Benaroya Research Institute

Steven G. Deeks sdeeks@php.ucsf.edu university of California San Fransisco

Giuseppe Del Giudice giuseppe.del_giudice@novartis.com Novartis

Martine Denis martine.denis@sanofipasteur.com Sanofi pasteur

Deborah Dunn-Walters deborah.dunn-walters@kcl.ac.uk Immunology, Infection And Inflammatory Disease King’s College london School of Medicine

Wendy Fantl wjfantl@stanford.edu Stanford university, Department of Microbiology and Immunology Jörg Goronzy jgoronzy@stanford.edu Stanford university School of Medicine

Speakers Teresa Aguado aguadom@who.int WHo Arne Akbar a.akbar@ucl.ac.uk university College london

Paolo Bonanni paolo.bonanni@unifi.it university of Florence, Department of public Health

Scott Boyd sboyd1@stanford.edu university of Stanford, Department of Pathology

Damien Chaussabel DChaussabel@benaroyaresearch.org Benaroya Research Institute

Steven G. Deeks sdeeks@php.ucsf.edu university of California San Fransisco

Giuseppe Del Giudice giuseppe.del_giudice@novartis.com Novartis

Martine Denis martine.denis@sanofipasteur.com Sanofi pasteur

Deborah Dunn-Walters deborah.dunn-walters@kcl.ac.uk Immunology, Infection And Inflammatory Disease King’s College london School of Medicine

Wendy Fantl wjfantl@stanford.edu Stanford university, Department of Microbiology and Immunology Jörg Goronzy jgoronzy@stanford.edu Stanford university School of Medicine

Harry Greenberg harry.greenberg@stanford.edu Stanford university, School of medicine

beatrix Grubeck-loebenstein beatrix.grubeck-loebenstein@oeaw.ac.at Institute For biomedical Aging Research of The Austrian Academy of Sciences

Agnes Hoffenbach Agnes.Hoffenbach@sanofipasteur.com Sanofi pasteur

Karl-Heinz Krause Karl-Heinz.Krause@unige.ch Dept. of pathology and Immunology, Geneva Medical Faculty Dept. of Genetic and laboratory Medicine, Geneva university Hospitals Myron levin Myron.levin@ucdenver.edu university of Maryland Gary Nabel gnabel@mail.nih.gov NIH

Janko Nikolich-Zugich nikolich@email.arizona.edu university of Arizona College of Medicine

Bali Pulendran bpulend@emory.edu university of emory Rino Rappuoli rino.rappuoli@novartis.com Novartis

Federica Sallusto federica.sallusto@irb.usi.ch Institute For Research In biomedicine Cellular Immunology Group leadervia

Albert Shaw albert.shaw@yale.edu yale university School of Medicine

This meeting was made possible through unrestricted grants from Novartis, Sanofi pasteur, Merck, Sanofi pasteur MSD.