Tissue Distributionof Human Y$ T Cells: No
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1012 J Clin Pathol 1991;44:1012-1017 Tissue distribution of human y$ T cells: No evidence for general epithelial tropism J Clin Pathol: first published as 10.1136/jcp.44.12.1012 on 1 December 1991. Downloaded from T M Vroom, G Scholte, F Ossendorp, J Borst Abstract peripheral lymphoid organs."6 More than In man and mice only a small proportion 90% of T cells in the murine epidermis, of T cells in the peripheral lymphoid however, express TCR y6,78 while y6 T cells compartment express the y8T cell recep- have also been reported to constitute a very tor (TCR). In mice, however, yB T cells large proportion of T cells in the epithelia of comprise the predominant population at reproductive organs,9 tongue,9 and intestine.'" particular epithelial sites-in epidermis Strikingly, certain Vy and V6 gene segments and epithelia of intestine, reproductive are expressed predominantly in different organs, and tongue. The distribution of murine tissues. Moreover, the diversity within y5 T cells in normal human tissues was the y6 T cell population differs greatly, depen- investigated, paying particular attention ding on the tissue. The epidermal y6 T cells to epithelial layers. In all lymphatic express V5/J1/Cy1 and VI/D2/J2/CQ encoded organs and in epithelia of a wide variety receptor chains" and can be considered clonal, of non-lymphatic organs, including the likewise the y3 T cell population in repro- respiratory tract, male and female re- ductive organs and tongue that expresses productive organs and tongue, y5 T cells V6/J1/Cy1 and V1/D2/J2/C6 encoded recep- constituted less than 5% of total T cells, tor chains.912 In intestinal epithelium" '3 and with the remainder expressing TCR 4p. in peripheral lymphoid organs'4 a large TCR The only exception was the intestine, yc repertoire exists that mainly depends on where yJ T cells were preferentially junctional diversity, while Vy7 and Vy4 gene situated in the columnar epithelium of segments are used preferentially at these re- the crypts, rather than in the lamina spective sites. Due to their specific localisation propria. in certain murine epithelia, their limited It is concluded, therefore, that human receptor diversity at two of these sites, and y5 T cells do not display a general epi- their potential specificity for the evolutionary thelial tropism and are, in terms of conserved heat shock proteins,"5 it has been relative numbers, no more able than a4 postulated that y6 T cells may have a special- T cells to out ised role in the carry continuous surveil- continuous immunosurveil- http://jcp.bmj.com/ lance of the immune system against in- lance of epithelia.""'8 fection or transformation in epithelia. yv In man TCR y6 bearing cells constitute less T cells may, however, have a specialised than 2% of CD3 positive thymocytes and less function in the epithelium of the intes- than 1-20% of peripheral blood T cells'920 In tinal tract. lymphoid organs y6 T cells constitute less than 5% of the T lymphocytes.202' With res- pect to the epithelial localisation of human y6 on October 1, 2021 by guest. Protected copyright. Two types of T cell antigen receptors (TCRs) T cells one clear difference with the murine are now known, the a, and the yb system is already apparent: in human skin y6 Department of heterodimer, which are expressed at the cell T cells are not the predominant T cell popula- Pathology, Rotterdam surface in association with the CD3 molecular tion, nor in epidermis nor dermis.2022 In intes- Cancer Center, complex.'` Each TCR chain contains a vari- tine the situation seems more comparable be- Rotterdam, The Netherlands able and a constant domain, encoded by dif- tween man and mice, with human yc T cells T M Vroom ferent gene segments V, (D), J and C that localising preferentially in the epithelium Department of combine by rearrangement during T cell dif- rather than in the lamina propria.2'2' Pathology, Slotervaart ferentiation. Although the y and loci We have quantitated y6 and ac, T cells in a Hospital, Amsterdam together contain fewer different of T M Vroom gene segments great variety normal human lymphatic and G Scholte than the a and loci, the potential repertoire non-lymphatic tissues, including the res- Division of of TCR yb is very large as one gene can piratory system, the male and female uro- Immunology, The incorporate more than one D segment,4 genital tract, and the tongue, which had not Netherlands Cancer which, with the addition of nucleotides at the previously been investigated. Special atten- Institute, Amsterdam F Ossendorp junctions, gives rise to a great variety of tion was paid to the various epithelial layers J Borst sequences.' At present, it is not clear what within these tissues to shed light on the pos- Correspondence to: contribution yb T lymphocytes make to the sible function of human yv T cells in epithelial Jannie Borst, Division of immune system. They may complement T surveillance. Immunology, The a# Netherlands Cancer cells in terms of antigenic specificities, func- Institute, Plesmnanan 121, tional capabilities, or sites of action within the 1066 CX Amsterdam, The Netherlands. body. Methods Accepted for publication In mice, yb T cells form a minority of the Normal tissues were obtained from necropsies 12 June 1991 total T cell population in the thymus and or from normal parts of surgical specimens. Tissue distribution ofhuman yv T cells 1013 Samples were derived from adults, unless was stained with haematoxylin and eosin for otherwise indicated. For every type of tissue histological examination. at least two samples from different subjects Monoclonal antibodies used for staining tis- were investigated. The following tissues were sues were: Identi-T #F1 (anti-TCR a)27 from J Clin Pathol: first published as 10.1136/jcp.44.12.1012 on 1 December 1991. Downloaded from examined: (1) lymphatic and haemopoietic T cell Sciences, Cambridge, Massachusetts, organs (a) thymus, fetal (12 weeks pregnancy) used at 2 Mg/ml; anti-TCR yb-l (hybridoma and neonatal, (b) lymph node, neonatal and name 1 F2, anti-TCR yb),'9 used at 2-4 adult, (c) tonsil, infantile, (d) spleen, (e) bone Mg/ml; CLB-T3 (anti-CD3) from Dr R van marrow, (f) liver, fetal (12 weeks pregnancy), Lier, Central Laboratory of the Red Cross neonatal and adult; (2) skin, fetal and adult; Blood Transfusion Service, Amsterdam, The (3) digestive tract (a) tongue, (b) salivary Netherlands, used at 5 pg/ml. Staining with glands, (c) oesophagus, (d) stomach, (e) small anti-TCR monoclonal antibodies was done intestine, (f) large intestine, (g) appendix; (4) according to the alkaline phosphatase-anti- urogenital tract (a) kidney, (b) ureter, (c) alkaline phosphatase method, with other urethra, (d) vagina, (e) uterus and cervix, (f) monoclonal antibodies according to the alka- testis and epididymis; (5) respiratory tract (a) line phosphatase method. Anti-TCR mono- nasal cavity, (b) trachea, (c) lung. clonal antibodies were tested for reactivity on Small tissue blocks were snap-frozen and cytocentrifuge preparations of TCR a,B stored in liquid nitrogen. Sections (8 gm positive and TCR yb positive T cell clones.'9 thick) were cut on a Reichert-Jung 2800 Frigocut Cryostat (Reichert-Jung GmbH, Nussloch, Germany), air dried, and fixed in acetone for 10 minutes. Fixation and all sub- Results sequent washes and incubations were perfor- To determine the proportion ofT lymphocytes med at room temperature. After fixation, sec- that expressed either TCR axf or TCR yb, serial tions were washed three times in phosphate sections were stained with anti-TCR 4a#, anti- buffered saline (PBS), pH 7-2. For the im- CD3, and anti-TCR yb monoclonal antibodies, munoalkaline phosphatase detection method in this order. Only tissue sections that showed sections were incubated with monoclonal no evidence ofinflammation were included, but antibody, diluted in PBS with 1% bovine intestine, lung, and endometrium and vagina serum albumin (BSA) (PBS/BSA), washed always show more or less reactive lesions. three times in PBS and incubated with alka- line phosphatase conjugated rabbit anti- LYMPHATIC AND HAEMOPOIETIC ORGANS mouse immunoglobulin (RaMIg-AP, Dako- In neonatal and fetal thymus less than 5% of patts D314, Glostrup, Denmark), diluted 1 in CD3 positive cells expressed TCR y6. In 20 in PBS with 10% normal human serum. accordance with published data,20 yb T cells For the alkaline -phosphatase-anti-alkaline were found preferentially in the juxta- phosphatase detection method, sections were medullary region of the cortex and in the first incubated with undiluted normal rabbit medulla and rarely present in the outer cortex. serum, followed by incubation with mono- No preferential association of yb T cells with http://jcp.bmj.com/ clonal antibody and then unconjugated rabbit Hassall's corpuscles was found. anti-mouse immunoglobulin (Dako Z259), In lymph nodes, tonsil, and Peyer's patches diluted 1 in 25 in PBS/BSA with 10% normal less than 5% of CD3 positive cells expressed human serum, as second step reagent, fol- TCR yb. In contrast to a,B T cells, yb T cells lowed by washing with PBS and incubation were not found within the lymph follicles. Of with alkaline phosphatase-anti-alkaline phos- the numerous T cells present within the lym- phatase complex (Dako D65 1), diluted 1 in 40 pho-epithelial area of the tonsillar crypts, less on October 1, 2021 by guest. Protected copyright. in PBS/BSA. Subsequently, sections were than 5% expressed TCR yb. washed three times in TRIS-buffered saline, In spleen yb T cells were again less than 5% pH 7-6, and incubated with staining solution of total CD3 positive cells in the periarteriolar for 30 minutes in the dark.