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Decidual-trophoblast interactions: decidual lymphoid cell populations in basal and parietal decidua
Journal of Reproductive Immunology 28 (1995) 165- 171
Brief communication
Decidual-trophoblast interactions: decidual lymphoid cell populations in basal and parietal decidua Herman Haller”, Francesco Tedescoc, Daniel Rukavina* b, Oriano RadilloC, Lea Gudelj b, Alan E. Beer d ‘Clinic for Obstetrics and Gynecology, Medical Faculty, University of Rijeka. B, Branchetta 20/l, 51000 Rijeka, Croatia bDepartment Physiology and Immunology, Medical Faculty, University of Rijeka. B. Branchetta 20/l, 5RWO Rijeka, Croatia ‘Istituto per I’mfanzia, Trieste, Italy ‘Department Microbiology and Immunology, Finch University Health Sciences, Chicago Medical School, Chicago, IL USA
Accepted 25 October 1994
Abstract
Immunohistochemical analysis of tissue specimens from human pregnancy decidua basalis in contact with invasive trophoblast of chorion frondosum and decidua parietalis in contact with non-invasive chorion laeve do not differ in the frequency of lymphoid cells of the following phenotypes (CD2, CD4, CDS, CD14, CD21 and gamma/delta TCR). A practical implication of this observation is that the collection of lymphoid cells from whole decidua by curettage for functional studies is justified. Keywords: Decidual lymphocytes; Decidua basalis; Decidua parietalis
+ Corresponding
author.
0165-0378/95/%09.50 0 1995 Elsevier Science Ireland Ltd. All rights reserved SSDI 0165-0378(94)00913-R
166
H. Hailer et al. /J. Reprod. Immunol. 28 (199s) 165-171
Prior to and during pregnancy, the uterine epithelium and stroma undergo changes known as the decidual reaction. This involves the appearance of highly specialized cell classes which are end products of proliferation and differentiation of libroblast-like stromal cell precursors (Dallenbach-Helweg, 1981). Immigrant and resident leukocytes are also recognized components of endometrial and decidual stroma (Bulmer and Sunderland, 1984; Kabawat et al., 1985). Three principal populations of decidual leukocytes are recognized by immunohistological techniques and more recently by flow cytometric analysis (Starkey et al., 1988; Chemishov et al., 1993). These are decidual macrophages (Bulmer and Johnson, 1984; Kabawat et al., 1985), large granulated lymphocytes of the natural killer cell lineage, most of which bear CD2 and CD56 antigens, but are negative for CD3, CD4, CD8 and CD16, and T lymphocytes (Bulmer and Sunderland, 1983, 1984; Bulmer et al., 1987, 1991, Haller et al., 1993). A definition of these cell types, numbers and immunoregulatory functions in normal and pathological pregnancies is the subject of other reports (Haller et al., 1993; Petrovid et al., 1994). The extent to which the patterns of lymphoid cells observed in decidual tissue obtained by curettage at the time of pregnancy terminations resembles that in vivo in decidua basalis as compared to decidual parietalis is unknown. Decidual tissue shows marked regional differences (basalis, parietalis, capsularis) in man and animals. The focus of immunological studies has centered primarily on the decidua basalis. As has been pointed out by Beer (1988), the decidua parietalis covers a vast expanse of fetal membranes. Both of these frontiers within the uterus contain cells with morphologic features of antigen presenting lymphocytes and lymphocytes capable of immune reactivity that could be both beneficial and harmful to the fetoplacental unit (Beer et al., 1994). To address possible regional differences in decidual cell populations we conducted an immunohistochemical analysis of tissue samples of both basal and parietal decidua obtained under directed ultrasonic guidance of the type utilized for chorionic villus biopsy for genetic diagnosis. This approach represents a model system that allows accurate evaluation of regional differences morphologically. Although functional assays of isolated decidual lymphoid cells were planned, technical limitations of too low cell numbers in samples prevented these definitive studies. Immunophenotypic analysis by FACS as well as functional studies of isolated decidual cells from normal and pathological pregnancies terminated by curettage have been reported (Petrovic et al., 1994). The investigations reported were approved by the Ethics Committee of the Medical Faculty, University of Rijeka. Biopsy specimens of basal and parietal decidua were obtained under ultrasound guidance from six pregnant women undergoing an elective pregnancy termination of normally progress-
H. Halier et al. /J. Reprod. Immunol. 28 (1995) 165-171
167
ing pregnancies between 6 and 9 weeks of gestation. For biopsy of basal decidua the forceps tip was introduced under the thickest area of the trophoblast. Three biopsy samples of both basal decidua and parietal decidua were obtained. The biopsy samples measured 3 x 3 x 5 mm and were snap frozen in liquid nitrogen and stored at -70°C. For immunohistochemical studies, 6-pm sections of frozen tissue were cut using cryostat (Frigocut, Pabish, Milan, Italy), air-dried, fixed in acetone/chloroform (1: 1) for 10 min at room temperature and either used immediately or kept at -70°C wrapped in aluminium foil. Immunoperoxidase staining of the sections was carried out by the PAP method as described previously (Tedesco et al., 1990) using Ortho universal mouse kit (Ortho Diagnostic System, Milan, Italy) and the following monoclonal antibodies indicated according to their specificity and dilutions: CD4 (Leu 3a + 3b Becton Dickinson, 1:2), CD8 (OKT8 - Ortho, 1:50), CD2 (OKT 11 Ortho, l:SO), CD56 (NKH-1 - Coulter, 1:20), CD14 (Anti-macrophages Dako, 1:20), CD21 (OKB 7 - Ortho, 150) and Gamma/Delta TCR (Anti TCR delta 1 - T cell Sciences, 1:20). All the antibodies had been tested on the frozen sections of human tonsil as positive control, and showed normal reactivity regarding the number and distribution of cells expressing tested antigens. Positive cells were counted randomly at 320 x magnification in 15 fields. Only single positive cells in decidual stroma were counted, while those forming clusters (follicles) were omitted in order to avoid errors while counting. The exact percentage of cells belonging to the corresponding subpopulations can be obtained by analyzing cell suspension prepared by enzymatic digestion of decidual tissue (Rukavina et al., 1994). However, it is also of interest to analyze the distribution of decidual mononuclear cells in situ, as was done in the present investigation. The leucocyte patterns exhibited in the decidual stroma of basal and parietal decidua were the same. Macrophages (CD14+ cells) were the predominant cell population and they were found evenly distributed in all areas of decidual tissue except for the decidual follicles. CD56’ cells were the second most common leukocyte population encountered in human first trimester decidua. These cells were uniformly dispersed throughout the de&dual tissue. CD56’ cells formed clusters around spiral arteries and at sites adjacent to endometrial glands. The numbers of CD56’ cells in parietal decidua were higher than in basal decidua, but the difference was not significant. CD4+ cells and to a somewhat lesser extent CD8+ cells were numerous in stroma, but found in follicles as well. The numbers of the cells were similar in both decidua basalis and decidua parietalis. There was a difference in the CD4:CD8 ratio which is higher in decidua basalis (2.0) than in decidua parietalis (1.6), but the difference is not significant. Gamma/delta TCR+ cells and CD21 + B lymphocytes were rarely found. Table 1 gives a
168
H. Hailer et al. /J. Reprod. Immunol. 28 (1995) 165-171
Table 1 Immunohistochemical
study of first trimester human decidua (n = 6)
Antigens
Basal dsecidua”
Parietal decidua”
CD4
324 zt 165 f 195 f 486 f ++++
314 l 97 198 zt 83 188 ztz24 597 f 111 ++++ f f
CD8 CD2 8 CD56 CD14 CD21 Gamma/Delta TCR
l l
74 52 63 171
‘Mean number and standard deviation of positive cells in 15 random fields (320 x ). ++++, More than 600 positive cells in 15 fields and f, Occasionally found positive cells.
quantitative evaluation of cells present in the decidual stroma (cells within decidual follicles are not reported). The procedure of collecting biopsy specimens of human decidua under ultrasonic control employed in this study allows characterization of immunocompetent lymphoid cells localized both at the implantation site, decidua basalis and at more distant sites, decidua parietalis. Data collected from animal experiments suggest that decidual cellular organization is highly localized in some animal species and is different in various regions of the uterus, supporting normal pregnancy in one segment of the uterus and allowing fetal resorption and abortion in a different region of the same uterine horn (Beaman and Hoversland, 1988). While investigating the immunocompetence of the pregnant uterus in rats, Beer (1988) showed that skin allografts survived the duration of gestation when placed at the choriodecidual junctions of mothers mated with syngeneic or allogeneic males. By contrast, all allografts implanted on the pseudodecidua of the uterine horn opposite to the one bearing the conceptuses were densely infiltrated by leukocytes and rejected. The decidua juxtaposed to the trophoblast appears to be a different ‘organ’ immunologically from the uterine tissue in the non-pregnant but decidualized horn. The results of the present investigation indicate that decidua basalis, in contact with invasive trophoblast of chorion laeve in humans, does not differ from the decidua parietalis in regard to the frequency or phenotype of lymphoid cells residing in each compartment, at least not with the panel of monoclonal antibodies against lymphocyte differentiation antigens used in these experiments. Unfortunately cell yield from these biopsy specimens did not permit FACS phenotypic and functional assays as we have reported from curretage specimens of normal pregnancies undergoing elective terminations and pathological pregnancies - anembryonic, missed abortion and ectopic (Petrovic et al., 1994).
H. Hailer et al. /J. Reprod. Immunol. 28 (1995) 165-171
169
There are no differences in the frequency of immunocompetent cell populations (T lymphocytes, B lymphocytes, macrophages and NK cells) in decidua basalis when compared to decidua parietalis. The functions of NK cells in decidual compartments have received much attention in the literature (King et al., 1989; Christmas et al., 1990; King and Loke 1991; Johnson et al., 1993). These are trophoblast growth promoting, clearing abnormal placental cells and preventing vertical viral transmission to the fetus. There are profound differences in trophoblast growth and differentiation adjacent to decidua basalis when compared to decidua parietalis. The expression of HLA G and embryonic antigens on extravillous and chorionic trophoblast may be important in this regard (Kovats et al., 1991; Tamake, 1992; Chiang et al., 1994). Our results clearly show that there is no significant difference in NK cell numbers in decidua basalis when compared to parietalis, in fact the numbers of these cells in decidua parietalis is slightly higher. Because of this it is hard to assign trophoblast growth promoting functions as the major role for decidual NK cells. In the present investigation we have found very few $6 TCR+ T lymphocytes as reported also by Diet1 et al. (1990). However, Mincheva et al. (1992) claimed that T cells expressing y/S TCR were abundant in decidua. For explanation of differences observed it should be emphasized that TCR 6-l monoclonal antibody used in our investigations was inefficient in immunohistochemical staining of decidual y/6 T lymphocytes, as was also in experiments reported by Diet1 et al. (1990) and confirmed by Mincheva et al. (1992). By using a mixture of mAb CyMl (directed against a constant part of y chain) and mAb SVl, Mincheva et al. (1992) were able to show that in cell suspension of decidual lymphocytes there is approximately 27% of y/6 positive cells. In this investigation we observed a prevalence of CD4’ over CD8’ cells, which is in contradiction with the cited results by Mincheva et al. (1992). However, it should be born in mind that there is a difference in the percentages of cell subpopulations obtained by analyzing decidual cell suspensions and decidual biopsies, especially in the protocols like ours where only individual cells scattered throughout the stroma (not aggregates) were numbered. In other investigations (Rukavina et al., 1995, by analyzing cell suspensions, we obtained only slight prevalence of CD4’ vs. CDS+ cells. A practical implication of these observations are that decidua basalis and decidua parietalis share a similar cellular pattern, i.e. that the collection of lymphoid cells from whole decidua by curettage for functional studies in humans seems justified. It should none the less be borne in mind that the relative proportion of cells bearing various markers may not be the same in tissue sections as in the cell suspensions. This investigation was supported by the Ministry of Science and Technology of the Republic of Croatia (Project No. 3-01-164).
170
H. Hailer et al. /J. Reprod. Immunol. 28 (1995) 165-171
Beaman, K. and Hoversland, R. (1988) Induction of abortion with monoclonal antibody specific for suppressor T-lymphocyte molecules. J. Reprod. Fertil. 82, 691-696 Beer, A.E. (1988) Immunology of reproduction. In: Immunological Diseases (Samter, M., ed.), pp. 329-360. Little Brown and Co., Boston/Toronto. Beer, A.E., Kwak, J.Y.H. and Ruiz, J. (1994) Reproductive Immunology. In: Principles and Practice of Clinical Immunology (Fleisher, T.A., Schwartz, B.D. and Shearer, W.T., eds.), Mosby Publishers (in press). Bulmer, J.N., Hollings, D. and Ritson, A. (1987) Immunocytochemical evidence that endometrial stromal granulocytes are granulated lymphocytes. J. Pathol. 153, 281-288. Bulmer, J.N. and Johnson P.M. (1984) Macrophage populations in the human placenta and amniochorion. Clin. Exp. Immunol. 57, 393-403. Bulmer, J.N., Morison, L., Longfellow, M. and Ritson, A. (1991) Leukocytes in human decidua: investigation of surface markers and function. In: Cellular and Molecular Biology of the Materno-Fetal Relationship (Chaouat, G. and Mowbray, J., eds.), pp. 189-196. INSERMJ. Libbey Eurotext, Paris-London. Bulmer, J.N. and Sunderland, C.A. (1983) Bone-marrow origin of endometrial granulocytes in the early human placental bed. J Reprod. Immunol. 5, 383-387. Bulmer, J.N. and Sunderland, C.A. (1984) Immunohistological characterization of lymphoid cell populations in the early human placental bed. Immunology 52, 349-357. Chemishov, V.P., Slukvin, 1.1. and Bondarenko, G.I. (1993) Phenotypic characterization of CD7+, CD3+ and CDS+ lymphocytes from first trimester human decidua using two-color flow cytometry. Am. J. Reprod. Immunol. 29, 5-16. Christmas, S.E., Bulmer, J.N., Maeger, A. and Johnson, P.M. (1990) Phenotypic and functional analysis of human CD3- decidual leukocyte clones. Immunology 11, 182- 189. Chiang M.H., Colbem, G.T. and Main, E.K. (1994) Identification of HLA-G polymorphism using denaturing gradient gel electrophoresis (DGGE) followed by DNA sequence analysis. Serono Symposium Boston, MA (in press). Dallenbach-Hellweg, G. (1981) The normal histopathology of the endometrium. In: Histopathology of the Endometrium, pp. 22-88. Springer-Verlag, New York. Dietl, J., Horny, H.P., Ruck, P., Mutzusch, E., Kaiserling, E., Greisser, H. and Kabelitz, D. (1990). Intradecidual T lymphocytes lack immunohistochemically detectable T-cell receptor. Am. J. Reprod. Immunol. 24, 33 Haller, H. Radillo, O., Rukavina, D., Tedesco, F., Candussi, G., Petrovic, 0. and Randic, L.J. (1993). An immunohistochemical study of leukocytes in human endometrium, first and third trimester basal decidua. J. Reprod. Immunol. 23, 41-49. Johnson, P.M. Deniz, G. and Mclaughlin, P.J. (1993) Functional properties of cloned CD3decidual leukocytes and low-affinity cytokine receptor expression on human trophoblast. In: Reproductive Immunology (Dondero, F. and Johnson, P.M., eds.), Vol. 97, pp, 141-144. Raven press, Rome. Kabawat, S.E., Mostoufi-Zadeh, M., Driscoll, S.G. and Bhan, A.K. (1985) Implantation site in normal pregnancy. A study with monoclonal antibodies. Am. J. Pathol. 118, 76-84. King, A. and Loke, Y.W. (1991). On the nature and function of human uterine granular lymphocytes. Immunol. Today. 12, 432-435. King, A., Birkby, C. and Loke, Y.W. (1989) Early human decidual cells exhibit NK activity against the K562 cell line but not against first trimester trophoblast. Cell. Immunol. 118, 337-344.
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Kovats, S., Librach, C., Fisch, P., Main, E.K., Sondel, P.M., Fischer, S.J. and DeMars, R. (1991) Expression and possible function of the HLA-G alpha chain in human cytotrophoblasts. In: Cellular and Molecular Biology of the Matemo-Fetal Relationship (Chaouat, G. and Mowbray, J. eds.), pp. 212-219. Colloque INSERM/John Libbey Eurotext Ltd. Mincheva-Nielson, L., Hammarstrom, S. and Hammarstrom, M.L. (1992) Human decidual leukocytes from early pregnancy contain high numbers of rS+ cells and show selective down - regulation of alloreactivity. J. Immunol. 149, 2203-2211 Petrovic, O., Gudelj, L., Rubesa, G., Haller, H., Beer, A.E. and Rukavina, D. (1995). Decidual-trophoblast interactions: Decidual lymphoid cell function in normal, anembrionic, missed abortion and ectopic human pregnancy. J. Reprod. Immunol. 26,217-23 1. Rukavina, D., Rubesa, G., Gudelj, L., Haller, H. and Podack, E.R. (1994) Characteristics of perforin expressing lymphocytes within the first trimester decidua of human pregnancy. Am. J. Reprod. Immunol. (in press) Starkey, P.M., Sarrgent, I.L. and Redman, G.W.G. (1988) Cell populations in human early pregnancy decidua: characterization and isolation of large granular lymphocytes by flow cytometry. Immunology 65, 129- 134. Tamake, J. (1992) Analysis of polymorphism at the human class 1 MHC locus, HLA-G. Hokkaido. Igaku. Zasshi. 67, 583-594. Tedesco, F., Radillo, O., Candussi, G., Nazzaro, A., Mollnes, T.E. and Pecorari, D. (1990) Immunohistochemical detection of terminal complement complex and S-protein in normal and preeclamptic placentae. Clin. Exp. Immunol. 80, 236-240.
Brief communication
Decidual-trophoblast interactions: decidual lymphoid cell populations in basal and parietal decidua Herman Haller”, Francesco Tedescoc, Daniel Rukavina* b, Oriano RadilloC, Lea Gudelj b, Alan E. Beer d ‘Clinic for Obstetrics and Gynecology, Medical Faculty, University of Rijeka. B, Branchetta 20/l, 51000 Rijeka, Croatia bDepartment Physiology and Immunology, Medical Faculty, University of Rijeka. B. Branchetta 20/l, 5RWO Rijeka, Croatia ‘Istituto per I’mfanzia, Trieste, Italy ‘Department Microbiology and Immunology, Finch University Health Sciences, Chicago Medical School, Chicago, IL USA
Accepted 25 October 1994
Abstract
Immunohistochemical analysis of tissue specimens from human pregnancy decidua basalis in contact with invasive trophoblast of chorion frondosum and decidua parietalis in contact with non-invasive chorion laeve do not differ in the frequency of lymphoid cells of the following phenotypes (CD2, CD4, CDS, CD14, CD21 and gamma/delta TCR). A practical implication of this observation is that the collection of lymphoid cells from whole decidua by curettage for functional studies is justified. Keywords: Decidual lymphocytes; Decidua basalis; Decidua parietalis
+ Corresponding
author.
0165-0378/95/%09.50 0 1995 Elsevier Science Ireland Ltd. All rights reserved SSDI 0165-0378(94)00913-R
166
H. Hailer et al. /J. Reprod. Immunol. 28 (199s) 165-171
Prior to and during pregnancy, the uterine epithelium and stroma undergo changes known as the decidual reaction. This involves the appearance of highly specialized cell classes which are end products of proliferation and differentiation of libroblast-like stromal cell precursors (Dallenbach-Helweg, 1981). Immigrant and resident leukocytes are also recognized components of endometrial and decidual stroma (Bulmer and Sunderland, 1984; Kabawat et al., 1985). Three principal populations of decidual leukocytes are recognized by immunohistological techniques and more recently by flow cytometric analysis (Starkey et al., 1988; Chemishov et al., 1993). These are decidual macrophages (Bulmer and Johnson, 1984; Kabawat et al., 1985), large granulated lymphocytes of the natural killer cell lineage, most of which bear CD2 and CD56 antigens, but are negative for CD3, CD4, CD8 and CD16, and T lymphocytes (Bulmer and Sunderland, 1983, 1984; Bulmer et al., 1987, 1991, Haller et al., 1993). A definition of these cell types, numbers and immunoregulatory functions in normal and pathological pregnancies is the subject of other reports (Haller et al., 1993; Petrovid et al., 1994). The extent to which the patterns of lymphoid cells observed in decidual tissue obtained by curettage at the time of pregnancy terminations resembles that in vivo in decidua basalis as compared to decidual parietalis is unknown. Decidual tissue shows marked regional differences (basalis, parietalis, capsularis) in man and animals. The focus of immunological studies has centered primarily on the decidua basalis. As has been pointed out by Beer (1988), the decidua parietalis covers a vast expanse of fetal membranes. Both of these frontiers within the uterus contain cells with morphologic features of antigen presenting lymphocytes and lymphocytes capable of immune reactivity that could be both beneficial and harmful to the fetoplacental unit (Beer et al., 1994). To address possible regional differences in decidual cell populations we conducted an immunohistochemical analysis of tissue samples of both basal and parietal decidua obtained under directed ultrasonic guidance of the type utilized for chorionic villus biopsy for genetic diagnosis. This approach represents a model system that allows accurate evaluation of regional differences morphologically. Although functional assays of isolated decidual lymphoid cells were planned, technical limitations of too low cell numbers in samples prevented these definitive studies. Immunophenotypic analysis by FACS as well as functional studies of isolated decidual cells from normal and pathological pregnancies terminated by curettage have been reported (Petrovic et al., 1994). The investigations reported were approved by the Ethics Committee of the Medical Faculty, University of Rijeka. Biopsy specimens of basal and parietal decidua were obtained under ultrasound guidance from six pregnant women undergoing an elective pregnancy termination of normally progress-
H. Halier et al. /J. Reprod. Immunol. 28 (1995) 165-171
167
ing pregnancies between 6 and 9 weeks of gestation. For biopsy of basal decidua the forceps tip was introduced under the thickest area of the trophoblast. Three biopsy samples of both basal decidua and parietal decidua were obtained. The biopsy samples measured 3 x 3 x 5 mm and were snap frozen in liquid nitrogen and stored at -70°C. For immunohistochemical studies, 6-pm sections of frozen tissue were cut using cryostat (Frigocut, Pabish, Milan, Italy), air-dried, fixed in acetone/chloroform (1: 1) for 10 min at room temperature and either used immediately or kept at -70°C wrapped in aluminium foil. Immunoperoxidase staining of the sections was carried out by the PAP method as described previously (Tedesco et al., 1990) using Ortho universal mouse kit (Ortho Diagnostic System, Milan, Italy) and the following monoclonal antibodies indicated according to their specificity and dilutions: CD4 (Leu 3a + 3b Becton Dickinson, 1:2), CD8 (OKT8 - Ortho, 1:50), CD2 (OKT 11 Ortho, l:SO), CD56 (NKH-1 - Coulter, 1:20), CD14 (Anti-macrophages Dako, 1:20), CD21 (OKB 7 - Ortho, 150) and Gamma/Delta TCR (Anti TCR delta 1 - T cell Sciences, 1:20). All the antibodies had been tested on the frozen sections of human tonsil as positive control, and showed normal reactivity regarding the number and distribution of cells expressing tested antigens. Positive cells were counted randomly at 320 x magnification in 15 fields. Only single positive cells in decidual stroma were counted, while those forming clusters (follicles) were omitted in order to avoid errors while counting. The exact percentage of cells belonging to the corresponding subpopulations can be obtained by analyzing cell suspension prepared by enzymatic digestion of decidual tissue (Rukavina et al., 1994). However, it is also of interest to analyze the distribution of decidual mononuclear cells in situ, as was done in the present investigation. The leucocyte patterns exhibited in the decidual stroma of basal and parietal decidua were the same. Macrophages (CD14+ cells) were the predominant cell population and they were found evenly distributed in all areas of decidual tissue except for the decidual follicles. CD56’ cells were the second most common leukocyte population encountered in human first trimester decidua. These cells were uniformly dispersed throughout the de&dual tissue. CD56’ cells formed clusters around spiral arteries and at sites adjacent to endometrial glands. The numbers of CD56’ cells in parietal decidua were higher than in basal decidua, but the difference was not significant. CD4+ cells and to a somewhat lesser extent CD8+ cells were numerous in stroma, but found in follicles as well. The numbers of the cells were similar in both decidua basalis and decidua parietalis. There was a difference in the CD4:CD8 ratio which is higher in decidua basalis (2.0) than in decidua parietalis (1.6), but the difference is not significant. Gamma/delta TCR+ cells and CD21 + B lymphocytes were rarely found. Table 1 gives a
168
H. Hailer et al. /J. Reprod. Immunol. 28 (1995) 165-171
Table 1 Immunohistochemical
study of first trimester human decidua (n = 6)
Antigens
Basal dsecidua”
Parietal decidua”
CD4
324 zt 165 f 195 f 486 f ++++
314 l 97 198 zt 83 188 ztz24 597 f 111 ++++ f f
CD8 CD2 8 CD56 CD14 CD21 Gamma/Delta TCR
l l
74 52 63 171
‘Mean number and standard deviation of positive cells in 15 random fields (320 x ). ++++, More than 600 positive cells in 15 fields and f, Occasionally found positive cells.
quantitative evaluation of cells present in the decidual stroma (cells within decidual follicles are not reported). The procedure of collecting biopsy specimens of human decidua under ultrasonic control employed in this study allows characterization of immunocompetent lymphoid cells localized both at the implantation site, decidua basalis and at more distant sites, decidua parietalis. Data collected from animal experiments suggest that decidual cellular organization is highly localized in some animal species and is different in various regions of the uterus, supporting normal pregnancy in one segment of the uterus and allowing fetal resorption and abortion in a different region of the same uterine horn (Beaman and Hoversland, 1988). While investigating the immunocompetence of the pregnant uterus in rats, Beer (1988) showed that skin allografts survived the duration of gestation when placed at the choriodecidual junctions of mothers mated with syngeneic or allogeneic males. By contrast, all allografts implanted on the pseudodecidua of the uterine horn opposite to the one bearing the conceptuses were densely infiltrated by leukocytes and rejected. The decidua juxtaposed to the trophoblast appears to be a different ‘organ’ immunologically from the uterine tissue in the non-pregnant but decidualized horn. The results of the present investigation indicate that decidua basalis, in contact with invasive trophoblast of chorion laeve in humans, does not differ from the decidua parietalis in regard to the frequency or phenotype of lymphoid cells residing in each compartment, at least not with the panel of monoclonal antibodies against lymphocyte differentiation antigens used in these experiments. Unfortunately cell yield from these biopsy specimens did not permit FACS phenotypic and functional assays as we have reported from curretage specimens of normal pregnancies undergoing elective terminations and pathological pregnancies - anembryonic, missed abortion and ectopic (Petrovic et al., 1994).
H. Hailer et al. /J. Reprod. Immunol. 28 (1995) 165-171
169
There are no differences in the frequency of immunocompetent cell populations (T lymphocytes, B lymphocytes, macrophages and NK cells) in decidua basalis when compared to decidua parietalis. The functions of NK cells in decidual compartments have received much attention in the literature (King et al., 1989; Christmas et al., 1990; King and Loke 1991; Johnson et al., 1993). These are trophoblast growth promoting, clearing abnormal placental cells and preventing vertical viral transmission to the fetus. There are profound differences in trophoblast growth and differentiation adjacent to decidua basalis when compared to decidua parietalis. The expression of HLA G and embryonic antigens on extravillous and chorionic trophoblast may be important in this regard (Kovats et al., 1991; Tamake, 1992; Chiang et al., 1994). Our results clearly show that there is no significant difference in NK cell numbers in decidua basalis when compared to parietalis, in fact the numbers of these cells in decidua parietalis is slightly higher. Because of this it is hard to assign trophoblast growth promoting functions as the major role for decidual NK cells. In the present investigation we have found very few $6 TCR+ T lymphocytes as reported also by Diet1 et al. (1990). However, Mincheva et al. (1992) claimed that T cells expressing y/S TCR were abundant in decidua. For explanation of differences observed it should be emphasized that TCR 6-l monoclonal antibody used in our investigations was inefficient in immunohistochemical staining of decidual y/6 T lymphocytes, as was also in experiments reported by Diet1 et al. (1990) and confirmed by Mincheva et al. (1992). By using a mixture of mAb CyMl (directed against a constant part of y chain) and mAb SVl, Mincheva et al. (1992) were able to show that in cell suspension of decidual lymphocytes there is approximately 27% of y/6 positive cells. In this investigation we observed a prevalence of CD4’ over CD8’ cells, which is in contradiction with the cited results by Mincheva et al. (1992). However, it should be born in mind that there is a difference in the percentages of cell subpopulations obtained by analyzing decidual cell suspensions and decidual biopsies, especially in the protocols like ours where only individual cells scattered throughout the stroma (not aggregates) were numbered. In other investigations (Rukavina et al., 1995, by analyzing cell suspensions, we obtained only slight prevalence of CD4’ vs. CDS+ cells. A practical implication of these observations are that decidua basalis and decidua parietalis share a similar cellular pattern, i.e. that the collection of lymphoid cells from whole decidua by curettage for functional studies in humans seems justified. It should none the less be borne in mind that the relative proportion of cells bearing various markers may not be the same in tissue sections as in the cell suspensions. This investigation was supported by the Ministry of Science and Technology of the Republic of Croatia (Project No. 3-01-164).
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