J Clin Pathol 1989;42:449-457

Occasional articles

Review

Use of basement membrane markers in tumour diagnosis

A J D'ARDENNE

From the Department ofHistopathology, St Bartholomew's Hospital, West Smithfield, London

Introduction

Basement membranes are complex structures com-posed of a mixture of collagenous and non-colla-genous glycoproteins and proteoglycans.'-5 Theirmolecular composition and supramolecular arrange-ment differ in different sites according to functionalrequirements. Basement membranes of epithelia andendothelia can be visualised as continuous limitingplates or tubes separating different body compart-ments. In the vascular system basement membranecontinuity is interrupted at only a few sites, notably thesinusoids of spleen and liver. Basement membranecomponents in splenic sinusoids are the structurallyspecialised ring fibres. In hepatic sinusoids only smallamounts of basement membrane material are presentin the space of Disse; no organised basement mem-brane is present. Elsewhere in the vascular systemspecialised filtering and diffusion properties are con-ferred by fusion of endothelial basement membraneswith those of adjacent epithelia. This occurs in renalglomeruli and pulmonary alveoli. Similar fusionoccurs in capillary basement membranes of the centralnervous system.'3

Basement membranes in mesenchymal tissues haveanother type of arrangement. In smooth muscle andadipose tissue individual cells are enveloped inbasement membrane. Schwann cells in nerves aresimilarly wrapped, providing continuous tubesthrough which the nerve fibres course."' Individualfibres of striated muscle are likewise surrounded by abasement membrane sheath. Notably, fibroblasts, thepotentially mobile cells of mesenchyme, do not have abasement membrane. The mobile cell populations oflymphoreticular and haemopoietic systems and cells incartilage and bone also lack this structure.

Several studies have indicated the importance ofbasement membranes in orderly tissue regenerationAccepted for publication 12 January 1989

and repair.' For example, an intact basement mem-brane is necessary for regeneration of renal tubularepithelium following acute tubular necrosis and forrepair of epidermis after skin damage. In the peri-pheral nervous system nerves can regenerate along thebasement membrane tubes in which they are normallyenveloped; neurons in the central nervous system donot have this capacity. Basement membranes are alsoimportant in embryogenesis for orderly cellmigration.5 Interactions between cells and basementmembranes occur, at least in part, via specific cellsurface receptors for different basement membraneconstituents. These extracellular proteins influence cellmorphology and differentiation as well as promotingcellular adhesion and movement.>"

Ultrastructurally, basement membranes arecharacterised by the presence of basal lamina. This isfurther subdivided into lamina densa and laminalucida. The lamina densa, sometimes-referred to as thebasement membrane proper, consists of tightly mattedrandomly orientated fibrils 3-4 nm in diameter,embedded in a dense matrix'; lamina lucida is anelectron lucent zone between plasma membranes andlamina densa, sometimes traversed by small filaments(figure).' In sites where fusion of two basementmembranes has occurred lamina lucida is present onboth sides of lamina densa-for example, in the renalglomerulus that on the epithelial side is the "laminarara externa", and that on the endothelial side is the"lamina rara interna".

Conventional methods of staining basement mem-branes, such as periodic acid Schiff (PAS) andreticulin, may stain not only basal lamina but alsostructures external to this. For example, manybasement membranes have an outer fibrillar "extrin-sic" layer known as lamina reticularis (figure).5 Bylight microscopy it is not possible to resolve thesedifferent components. Immunohistological methodsshow that antibodies directed against constituents of

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~Lamiino lucidoL Basail laminaBasement

membrane Lamino reticularis

Figure Diagrammatic representation ofan epithelialbasement membrane.

any of these structures will give rise to light micro-scopic basement membrane delineation.

Extraction and identification of basal laminaproteins was initially hampered by the difficulty ofobtaining pure basal lamina. The unique basementmembrane collagen (type IV) was first extracted fromglomerular basement membranes that lack an outerlamina reticularis.'2 Purification of other basal laminaconstituents was facilitated by the culture of tumourcell lines producing basal lamina in excess, notably theEngelbreth-Holm-Swarm (EHS) sarcoma'3 andteratocarcinoma cells. 14

Principal constituents of basal lamina include typeIV collagen,'5 the high molecular weight glycoproteinlaminin,'6 and proteoglycans,'7 18 principally heparansulphate proteoglycan. Entactin'4 and nidogen'9 are

further basal lamina proteins. These are probablyrelated if not identical molecules.20 The adhesiveglycoprotein fibronectin is present in lamina reticularisas well as in some, if not all basal laminae, inassociation with cell surfaces.2' 24

The principal fibrillar components of laminareticularis are interstitial collagens types I and111.5 20 25 26 Additional collagenous molecules associatedwith some but not all basement membranes arecollagens types V, VI, and VII.213' Collagen type VII isprobably a principal constituent of anchoring fibrilswhich link the basal lamina of skin and other stratifiedepithelia to underlying connective tissue.32The heterogeneous composition of basement mem-

branes has been shown by production of monoclonalantibodies with selective reactivity for basement mem-branes in different sites.3334 In many cases the under-lying biochemical basis for this heterogeneity is notknown. It may be partly attributable to differingextrinsic components. Further evidence for heterogen-eity is provided by diseases such as Goodpasture'sdisease and pemphigoid, in which anti-basementmembrane autoantibodies are restricted to specificlocations.35 36

BASEMENT MEMBRANES IN TUMOURSA property fundamental to malignant disease isinvasiveness. It might seem that the first barrier a

carcinoma must transgress before infiltrating

d'Ardennesurrounding tissues is the basement membrane.3"Consequently many studies have investigated thispossibility. Early investigations used staining tech-niques such as PAS and reticulin.38 The advent of theelectron microscope permitted detailed ultrastructuralexamination of basal lamina in tumours and inflam-matory conditions.3"3 More recently, biochemicalcharacterisation and production of antibodies againstdifferent basement membrane constituents has meantthat application of immunohistological techniquescould be applied to the problem. This has the advan-tage over ultrastructural analysis that much largervolumes of tissue can be rapidly sampled andexamined.

Both ultrastructural and immunohistologicalstudies have shown that in general there is loss andfragmentation of basal lamina in malignant tumoursof both epithelial and mesenchymal origin.3"' Re-duplications may also be seen. Interpretation of thesephenomena has differed. It is clear that they reflectabnormal turnover ofbasement membranes due eitherto increased breakdown or decreased synthesis of theirconstituents.62' Less clear is the importance of thesechanges to tumour behaviour. They might representthe primary event in invasion.63 Alternatively, theymay simply be an epiphenomenon reflecting abnormalcellular differentiation.6719 The distinction has animportant bearing on the use of basement membranemarkers in tumour diagnosis.

Yet another interpretation of the importance ofbasement membrane constituents in neoplasia is thatsubstances such as laminin are actually necessary topromote invasion and metastasis.7° This conceptoriginated from experiments on transplantable murinetumours71: the ability of some tumours to metastasiseis related to their ability to secrete or bind tolaminin.7' An important factor may be whether theypossess unoccupied laminin receptors.7F76 These mighthelp tumour cells to bind to basement membranes ofvessels and those at distant sites, as well as to lamininproduced by the tumour cells themselves.The purpose of this review is to discuss whether

basement membrane immunohistological analysis canbe used to determine (a) if a tumour is invasive, (b) todistinguish malignant tumours from benign "look-alikes", (c) to determine tumour prognosis and (d) todetermine tumour histogenesis.

CHOICE OF MARKERFor immunohistological study of basal lamina it ispreferable to use an antibody directed against aprotein which is ubiquitous in this structure and whichis confined to this structure. It should also be possibleto show that immunohistological delineation of basallamina at light microscopy correlates with distributionof basal lamina observed ultrastructurally. Antibodies

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Use ofbasement membrane markers in tumour diagnosisagainst type IV collagen and against laminin for themost part seem to fulfil both these criteria. In tumours,however, it is sometimes possible to detect smallamounts of extracellular immunoreactive lamininwhere no basal lamina can be detected ultrastruc-turally." This may represent secreted protein whichhas not become organised into a macromolecularstructure. Alternatively, it may simply reflect thesampling problems inherent in ultrastructural examin-ation of tissue. It is also sometimes possible to showthe presence of intracellular laminin, presumablyreflecting synthetic activity of a cell."78

Immunohistological demonstration offibronectin isless suitable for investigating basal lamina changes perse as this protein is not confined to this structure.Observation of changed distributions of fibronectinand other basement membrane and stromal proteinsmay nevertheless prove of interest in their ownright.79

METHODSThe most reliable methods of showing the presence ofbasement membrane proteins in tissue sections entailthe use of fresh frozen tissue or tissue which has beenfixed in ethanol before processing in paraffin wax(Sainte Marie method).8' Type IV collagen, laminin,and fibronectin, however, can usually be effectivelyshown by immunohistological techniques in enzy-matically digested formalin fixed, paraffin wax embed-ded tissue.8286 This is an obvious advantage if fresh orspecially prepared tissue is unavailable. Digestion isessential and choice of enzyme is important. Forreasons that remain obscure, trypsin and some brandsof commercial protease are ineffective for use withthese extracellular proteins. Digestion with pepsin orprotease type VII or XXIV (Sigma) can produceoptimal results.86 Use of fixatives other than formalincan also affect staining. Whereas most polyclonalantisera against these proteins should allow them to beshown in routinely processed tissue, some monoclonalantibodies are unreactive in these conditions. Thismay be explained by loss ofimmunoreactivity of somebut not all epitopes of a protein during the fixation orembedding procedure.

Is it invasive?

Use of basement membrane immunohistological tech-niques to distinguish invasive from in situ neoplasiapresupposes that basement membrane disruption isessential for invasion to occur. The studies carried outby Barsky et al suggested that this, indeed, was thecase.5 The authors examined neoplasms from variousdifferent tissues including breast, skin, pancreas andprostate and reported that benign and in situ lesionshave intact basement membranes with linear staining

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for type IV collagen and laminin; most invasivecarcinomas lack immunoreactivity for both theseproteins. They reported that cases of in situ carcinomawith microinvasion show thinning, fragmentation,and disruption of the basement membrane in the fociof microinvasion but not elsewhere. Other studieshave been less absolute in their conclusions. Manyinvasive tumours show variable retention of basallamina proteins at their periphery.'58676987 Further-more, dysplastic and in situ lesions in a variety of sitesmay have discontinuous or interrupted basementmembrane staining.4953 59 6 78 Consequently, loss ofbasement membrane continuity cannot be used as asimple criterion of invasion.

Several studies of invasive breast carcinoma havedescribed variation in laminin immunoreactivity bothwithin and between tumours.44"4""s This ranges fromdiscontinuous and linear in the better differentiated,invasive ductal carcinomas to complete absence inmoderately differentiated and undifferentiatedtumours. Variation in quantity of laminin shown indifferent studies may reflect technical variations, inparticular, fixation. It may also reflect variation ininterpretation of the results as laminin in tumours maybe visualised as a much thinner and finer line than thatseen around normal glands.77 Notably, continuouslinear staining for type IV collagen and laminin, suchas that found around normal breast acini, has not beendescribed at the margins of invasive ductal and lobularbreast carcinoma. Fragmentation of basement mem-brane, however, may be seen around intraductalcarcinoma of breast."78

In contrast to breast carcinomas, linear type IVcollagen and laminin may be found at the margins ofwell differentiated squamous cell carcinomas of thelarynx, oropharynx, skin and cervix.' 58 676888 Invasivecarcinomas, however, generally show at least focaldiscontinuities of basement membrane and it may betotally lost. Basal cell carcinomas are usually com-pletely enveloped by basal lamina, although this is notinvariable and it tends to be fragmented in deeplyinfiltrative tumours.87 89 90

It has been suggested that the differing amount ofbasal lamina seen around different tumour types is areflection of their cell of origin and degree of differen-tiation.47 67 6 Relative paucity of basal lamina ininvasive breast carcinoma might be a reflection ofrelative paucity of myoepithelial differentiation inthese tumours. In contrast, squamous and basal cellcarcinomas might be expected to produce largeramounts. Other neoplasms producing large quantitiesof basal lamina are adenoid cystic carcinomas andsome pleomorphic adenomas.23 78 This would be con-sistent with their putative myoepithelial origin.Analogous to the breast, dysplastic and in situ

lesions ofsquamous epithelia of skin, oropharynx, and

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452larynx may be associated with either continuous orinterrupted basement membrane staining. Interrup-tions seem to be less common in dysplasia of thecervix, where linear immunoreactivity for the principalbasal lamina constituents has been reported.49 It wassuggested that this represents the longer time course ofprogression from in situ to invasive carcinoma in thissite and the possibility of regression49. In a series ofcases of cervical intraepithelial neoplasia, however,Richards and Furness described basement membraneinterruptions that increased in number with increasingseverity of dysplasia.9' Foci of microinvasive carcin-oma had a completely different pattern of immun-oreactivity, being absent or completely fragmented,and it was suggested that this could be of potentialdiagnostic use.

Although it is apparent that loss of basementmembrane integrity cannot in general be used as asimple diagnostic criterion of invasion, the abnor-malities of this structure associated with dysplasia andneoplasia demand further investigation. Diagnosticapplications depend on a better understanding ofchanges observed.

PROBABILITY OF INVASIONA particularly important question is whether loss ofbasement membrane integrity in dysplasia and in situcarcinoma indicates a greater probability of progres-sion to invasive malignancy. If this were proved to bethe case it might be a useful adjunct to gradingdysplasias and identifying the potentially most aggres-sive changes. This possibility has been explored in theurinary bladder.59 In a retrospective study of 69superficial and 15 invasive urothelial carcinomas itwas found that although superficial "non-invasive"tumours may have either intact or interrupted base-ment membranes, the incidence of progression ofsuperficial tumours with patchy or absent basementmembranes was significantly greater than those withcomplete basement membranes. There was someassociation between basement membrane staining andhistological grade but this did not reach significance.A complicating factor is that basement membrane

interruptions may be found in association with inflam-mation. It has been reported that breaks in basementmembrane at sites of inflammation are small andsharply defined in contrast to the irregular discontin-uities found in in situ carcinoma.' Others havesuggested that the gaps in basement membrane seen indysplasia may actually be due to enzymes produced byinflammatory cells and that this might promoteinvasion.59 Further studies are obviously required inthis area.

LIMITED TISSUEAnother situation in which it has been suggested that

d'Ardenneimmunostaining of the basal lamina may have adiagnostic role is in the analysis of endometrialcurettings. In a study of benign, premalignant, andmalignant endometrium Furness and Lam reportedthat in normal endometrium and benign cystic hyper-plasia, glandular basement membranes are nearlycontinuous even in the menstrual phase.57 Smallnumbers of breaks are found in atypical hyperplasia,with increasing numbers the greater the abnormality.Invasive tumours were said to have a strikinglydifferent pattern with many breaks in basement mem-brane, even when well differentiated. It was thoughtthat the differences between benign and atypicalhyperplasia were insufficient to be a useful diagnostictool. The differences between atypical hyperplasia andinvasive carcinoma, however, were thought to besufficiently distinctive to have a potential role in theanalysis of endometrial curettings where lack of tissuemakes invasion hard to assess.

In other studies of endometrium it has been notedthat stromal cells are enveloped by laminin but only atcertain stages of the menstrual cycle.9293 Stromallaminin was observed in secretory endometrium and in69% of cases of cystic hyperplasia. In contrast, it wasnot present in proliferative endometrium or in atypicalhyperplasia or carcinoma. This was also suggested tobe of potential diagnostic value.

TYPE VII COLLAGENA basement membrane antibody produced morerecently is the monoclonal antibody LH7 2 whichreacts with type VII collagen.9 This is only reactive onfresh, frozen tissue sections. Unlike the basal laminaantigens laminin and type IV collagen, type VIIcollagen is restricted to basement membranes ofstratified epithelia and is not present around naevuscells. In malignant melanomas it was found that loss ofan intact LH7-2 positive basement membrane beneathan intraepithelial proliferation of malignant melan-ocytes correlated with increased tumour thickness.95 Itwas suggested that the relatively good prognosis ofthin malignant melanomas may be associated withconfinement of the tumour above intact epidermalbasement membrane. The pattern of laminin and typeIV collagen deposition in malignant melanomas isanalagous to that in carcinomas with variable deposi-tion around tumour nests or individual tumour cellsand sometimes total absence.9 97The antibody to collagen type VII has also been

applied to basal cell carcinomas. Its pattern of reac-tivity was similar to that of laminin and type IVcollagen, but in general it was diminished in amountrelative to these two proteins and relative to strongstaining of epidermal basement membrane. In twodeeply infiltrative basal cell carcinomas fragmentationofbasal lamina staining was accompanied by total loss

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Use ofbasement membrane markers in tumour diagnosisof type VII collagen immunoreactivity (d'Ardenne et

al, unpublished observations). The presence or

absence of type VII collagen may have an influence onthe locally infiltrative capacity of these tumours.

Benign or malignant?

A slightly different diagnostic problem is the distinc-tion of an overtly invasive malignancy from a com-

pletely benign, non-neoplastic "look-alike". Twoexamples are the distinction of tubular carcinoma ofthe breast from sclerosing adenosis and distinction ofpancreatic adenocarcinoma from chronic pancreatitis.The possibility ofusing basement membrane immuno-histological techniques as a diagnostic aid has beenexplored in both these situations. Complete absence ofbasement membrane around tubular carcinomas hasbeen noted both ultrastructurally and by immunohis-tology.5 9 This is in contrast to continuous and linearstaining ofbasement membrane in benign lesions. Thismay be of diagnostic value provided the possibility offalse negative results is recognised (see below).The value of laminin staining for distinguishing

between chronic pancreatitis and adenocarcinoma ofthe pancreas has been investigated by Haglund et al.'They reported mostly irregular and discontinuouslaminin around invasive ductal carcinoma, although itwas almost continuous around some well differen-tiated glandular structures. They concluded thatimmunohistochemical analysis for laminin might be a

useful diagnostic aid provided enough tissue is availa-ble for examination. In contrast, it was not found to beuseful for distinguishing between benign and malig-nant mucinous cystic neoplasms or islet cell tumours,all of which possessed a nearly intact basementmembrane.

Its use has also been investigated in the thyroid todetermine if basement membrane staining might helpin the sometimes difficult distinction of encapsulated,well differentiated follicular carcinoma from follicularadenoma.' ' 02 It was found that basement membraneswere only consistently deficient in the widely invasivetumours-that is, those that do not present anydiagnostic difficulty. Encapsulated microinvasivetumours may have intact basement membrane aroundtrabeculae and follicles and this can be seen even inareas of vascular invasion. In some microinvasivetumours, however, there is widespread loss ofimmunoreactive laminin and fibronectin.A difficulty with the use of basal lamina markers to

distinguish benign from malignant neoplasms is thatthe positive diagnosis (malignancy), is supported bythe negative observation (lack of basal laminaimmunoreactivity). Extreme caution must thereforebe exercised to ensure that the immunostains haveactually "worked" if basal lamina markers are to beused in this manner. Positive staining of endothelial

453basement membranes is not always a reliable indicatorof staining efficacy as antigen accessibility may differin different types of basement membrane. For exam-ple, excessive deposition of interstitial collagen inepithelial basement membranes may mask basallamina antigens." Positive staining of non-neoplasticepithelial basement membranes in the test section isnevertheless the best internal positive control avail-able. In practice, this type of staining can only be usedto support diagnoses for which there is already strongsuspicion on other grounds.

Prognosis

Quantity of laminin around invasive carcinoma hasbeen reported to correlate with its degree of differen-tiation, but few formal studies of its relation toprognosis have yet been made. It might be predictedthat the better differentiated carcinomas would have agreater amount oflaminin and a better prognosis. Thismay prove a general rule but it may only apply to sometypes of neoplasm. Tubular and mucinous carcinomasof the breast are recognised to have a relatively goodprognosis and yet both are completely devoid of basallamina. As noted above, it also seems that the amountof basal lamina a tumour is likely to have dependspartly on its origin. Squamous and transitional cellcarcinomas tend to have more than adenocarcinomas,and adenocarcinomas of stomach and colon, forexample, tend to have more than adenocarcinomas ofthe breast. Each type of tumour therefore requiresseparate consideration.

In general, tumour prognosis is related to stage andgrade. Although there is a tendency for basal laminaquantity to be related to histological grade,69 103 thereseems to be a poor correlation between patterns ofbasal lamina and stage or extent ofspread ofa tumour.If basal lamina immunostaining is to be of value as aprognostic variable, it must provide further informa-tion to that already provided by conventional stagingand grading methods. Two studies have suggested thatthis is indeed the case. In a study of 75 bladdercarcinomas, 27 superficial and 48 invasive, Daher et alreported that the invasive tumours could be dividedinto two groups: those with conserved or only partiallyfragmented basal lamina; and those with widelyfragmented or absent basal lamina.'05 The latter hadsignificantly lower short term survival, independent ofstage or histological grade.

Forster et al described the results of lamininimmunostaining in a series of 158 rectal adenocarcin-omas.'06 Sixty two per cent had well defined basementmembrane laminin, and the corrected five yearsurvivals for laminin positive and laminin negativetumours were 65% and 23%, respectively. There waspartial correlation of laminin positivity with tumourgrade, but multivariate analysis indicated it is a better

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454 d'Ardenneindicator of prognosis than conventionally assessedhistological grade. Duke's staging remained the bestindependent prognostic variable.

Tumour histogenesis

Basement membrane markers cannot be used as"markers" of tumour cell histogenesis in the conven-tional sense as they are common to such a wide rangeof cell types. As described in the introduction,however, their organisation differs in different tissuesand a similar type of organisation may be found intheir malignant counterparts. When using basementmembrane markers in this context, it is important toremember that malignant tumours may lose theirbasement membranes altogether. A negative result istherefore non-contributory.

In attempting to differentiate a sarcoma from acarcinoma, the presence of basal lamina aroundindividual cells as opposed to groups of cells points toa sarcoma. This can occasionally be of value-forexample, in the diagnosis ofspindle cell tumours ofthekidney (d'Ardenne, unpublished observations). This isanalogous to use of a reticulin stain but the results aremore specific and easier to interpret. Presence of basallamina around individual tumour cells also indicatesthat they are not of fibroblastic origin as fibroblastslack this structure. Positive laminin staining in asarcoma excludes a diagnosis of fibrosarcoma or amalignant fibrous histiocytoma.6"'07 0 It may also behelpful in the diagnosis of vascular tumours toelucidate the relation of the tumour cells to basementmembranes, again in a manner analogous to a reticulinstain."09 "°A tumour which has a very distinctive pattern of

basal lamina deposition is adenoid cystic carcin-oma.4278"' In this neoplasm abundant basal laminamay be found either among individual tumour cells orlining the characteristic cystic spaces. This may be ofvalue in distinguishing adenoid cystic carcinomasfrom cribriform adenocarcinomas, especially in thebreast. In cribriform adenocarcinomas the cysticspaces represent true glandular lumina and are con-sequently not lined by basal lamina.

Demonstration of basal lamina is not of assistancein distinguishing adenoid cystic carcinomas frompleomorphic adenomas as both can have a very similarpattern of reactivity.23 In general, however, pleomor-phic adenomas have variable and irregular amounts ofbasal lamina at the margins of tumour islands.23 12

Identification of basal lamina has previously beenrecommended by ultrastructural pathologists as auseful landmark when attempting to solve the type ofproblem described above.' "' 1'3 The major advantageof using basal lamina immunohistological techniquesis the facility of looking at much larger volumes oftissue.

Conclusion

The relation of basal lamina to tumour invasiveness isa complex subject, many aspects of which have yet tobe clarified. Although it is apparent that invasioncannot be regarded simply as penetration ofneoplasticcells through a basement membrane barrier, the fewstudies done indicate that abnormalities of this struc-ture are found in most invasive tumours and in manysevere dysplasias. The importance ofthis phenomenonis indicated by the few studies in which basal laminaabnormalities have been related to subsequent tumourbehaviour. It is not possible with this type ofinvestiga-tion to determine whether basal lamina disruption issimply a manifestation of abnormal tumour differen-tiation. Nevertheless, as basal lamina is essential forthe maintenance of normal tissue architecture it seemslikely that abnormalities in basal lamina contribute tothe architectural disruption of neoplastic tissue. Thismay in turn be important for the occurrence of overtinvasion.From the diagnostic point of view it is clear that in

most situations delineation of basal lamina cannot beused as an absolute criterion to distinguish betweeninvasive and non-invasive malignancy. Possible excep-tions to this generalisation are a few situations inwhich the distinction to be made is between a totallybenign and an overtly malignant invasive tumour.An important question remaining to be fully ans-

wered is whether basal lamina status may provideadditional or possibly even more relevant informationabout invasive potential of dysplasias than commonlyapplied methods. Similarly, the prognostic importanceof basal lamina and its cell surface receptor moleculesin invasive neoplasms requires further investigation.A totally separate question is the value of basal

lamina markers in determining tumour histogenesis.In this situation their function is to supply architec-tural information as well as to distinguish betweenbasal lamina-producing and non-basal lamina-producing cells. The advent of monoclonal antibodieswith tissue restricted basement membrane reactivitymay enhance the specificity of this approach. Atpresent, basement membrane immunohistology canbe regarded as an occasionally helpful adjunct to otherdiagnostic methods.

AJd'A is supported in part by the Imperial CancerResearch Fund.

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