Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Unsuitability of the assay for cell-mediated lympholysis in inbred mice for H-Y antigen determination of human cells


This study examined the H-Y-specific in vitro restimulation of splenocytes from in vivo intraperitoneally (i.p.) primed C57B1/6 (B6) female mice. In vivo priming was carried out with human male or female fibroblasts or peripheral blood lymphocytes, respectively. It was attempted to measure the in vitro H-Y-specific activity by cell-mediated lympholysis and by cell proliferation. 3[H]Thymidine incorporation was determined in mixed lymphocyte cultures (MLCs) of xenogenetic primed female splenocytes (responder cells) and of syngeneic lethally irradiated male splenocytes (stimulator cells). The xenogenic H-Y presentation by in vivo sensitization did not induce in the in vitro restimulation system an H-Y-specific cell proliferation or in the effector phase the generation of H-Y-specific killer cells. The assay for celimediated lympholysis and lymphocyte proliferation after xenogeneic priming and syngeneic in vitro restimulation is, thus, not suitable for H-Y testing of human cells.

This is a preview of subscription content, log in to check access.


  1. Biasi G, Dazzi F, Loveland B, Rivarrollo R, Asherson G (1985) The failure of female cells to present in vitro the male H-Y antigen for secondary cytotoxic T-cell response. Immunogenetics 22:177–181

  2. Billingham RE, Silvers WK (1960) Studies on tolerance of the Y-chromosome antigen in mice. J Immunol 85:14–26

  3. Böhmer H von, Turton K, Haas W (1979) The role of the left end of the H-2b haplotype in the male-specific cytotoxic T cell response. Eur J Immunol 9:913–915

  4. Eichwald EJ, Silmser CR (1955) Untitled communication. Transplant Bull 2:148–149

  5. Fierz W, Brenan A, Müllbacher A, Simpson E (1982a) Non-H-2 and H-2-linked immune response genes control the cytotoxic T-cell response to H-Y. Immunogenetics 15:261–270

  6. Fierz W, Farmer GA, Sheena JH, Simpson E (1982b) Genetic analysis of the non-H-2-linked Ir genes controlling the cytotoxic T-cell response to H-Y in H-2d mice. Immunogenetics 16:593–601

  7. Gasser DL, Silver WK (1972) Genetics and immunology of sex-linked antigens. Adv Immunol 15:215–247

  8. Goldberg EH, Shen FW, Tokuda S (1973) Detection of H-Y (male) antigen on mouse lymph node cells by cell to cell cytotoxicity test. Transplantation 15:334–336

  9. Gordon RD, Simpson E, Samelson LE (1975) In vitro cell-mediated immune responses to the male specific (H-Y) antigen in mice. J Exp Med 142:1108–1120

  10. Gordon RD, Mathieson BJ, Samelson LE, Boyse EA, Simpson E (1976) The effect of allogeneic presentation on H-Y graft survival and in vitro cell-mediated responses to H-Y antigen. J Exp Med 144:810–820

  11. Goulmy E, Blokland E, van Rood J, Charmot D, Malissen B, Mawas C (1980) Production, expansion, and clonal analysis of T cells with specific HLA-restricted male lysis. J Exp Med 152:182–190

  12. Goulmy E, van Leeuween A, Blokland E, Sachs ES, Geraedts JPM (1983) The recognition of abnormal sex chromosome constitution by HLA restricted anti-H-Y cytotoxic T cells and antibody. Immunogenetics 17:523–531

  13. Hurme M, Hetherington CM, Chandler PR, Simpson E (1978a) Cytotoxic T-cell responses to H-Y: mapping of the Ir genes. J Exp Med 147:758–767

  14. Hurme M, Chandler PR, Hetherington CM, Simpson E (1978b) Cytotoxic T-cell responses to H-Y: correlation with the rejection of syngeneic male skin grafts. J Exp Med 147:768–775

  15. Juretié A, Malenica B, Juretié E, Klein J, Nagy ZA (1985) Helper effects required during in vivo priming for a cytologic response to the H-Y antigen in nonresponder mice. J Immunol 134:1408–1414

  16. Klein J (1986) How many class II immune response genes? A commentary on a reappraisal. Immunogenetics 23:309–310

  17. Liew FY, Simpson E (1980) Delayed-type hypersensitivity responses to H-Y: characterisation and mapping of the Ir genes. Immunogenetics 11:255–266

  18. Matsunaga T, Simpson E (1978) H-2 complementation in anti-H-Y cytotoxic T-cell responses can occur in chimeric mice. Proc Natl Acad Sci 75:6207–6210

  19. Müllbacher A, Brenan M (1980) Cytotoxic T-cell response to H-Y in “non-responder” CBA mice. Nature 285:34–36

  20. Pfeffer PF, Sodal G, Thorsby E (1981) HLA-restricted cell-mediated cytotoxicity directed against male specific (H-Y) antigen after acute rejection of an HLA-identical sibling kidney. Scand J Urol Nephrol [Suppl] 64:72–78

  21. Silvers WK, Yang SL (1973) Male specific antigen: homology in mice and rats. Science 181:570–572

  22. Simpson E, Gordon RD (1977) Responsiveness to H-Y antigen: Ir gene complementation and target cell specificity. Immunol Rev 35:59–75

  23. Simpson E, Liebermann R, Andó I, Sachs DH, Paul WE, Berzofsky JA (1986a) How many class II immune response genes? A reappraisal of the evidence. Immunogenetics 23:302–308

  24. Simpson E, Liebermann R, Andó I, Sachs DH, Paul WE, Berzofsky JA (1986b) A comment on the commentary. Immunogenetics 23: 311

  25. Waal LP de, Hoop de J, Stukart MJ, Gleichmann H, Melvold RW, Melief CJ (1983a) Nonresponsiveness to the male antigen H-Y in H-2 I-A-mutant B6.C-H-2bm 12 is not caused by defective antigen presentation. J Immunol 130:655–660

  26. Waal LP de, Melvold RW, Melief CJ (1983b) Cytotoxic T lymphocyte nonresponsiveness to the male antigen H-Y in the H-2Db mutants bm13 and bm14. Complementation of the response in F1 crosses with the I-Ab mutant bm12 nonresponder and failure of B6 or Db mutant mice tolerant of each other to respond to allogeneic male cells. J Exp Med 158:1537–1546

  27. Wiberg UH (1985) H-Y transplantation antigen in human XO females. Hum Genet 69:15–18

  28. Wiberg UH, Fredga K (1985) The H-Y transplantation antigen is present in XO and X*X female wood lemmings (Myopus schisticolor). Immunogenetics 22:495–501

  29. Wiberg UH, Günther E (1985) Female wood lemmings with the mutant X*-chromosome carry the H-Y transplantation antigen. Immunogenetics 21:91–96

  30. Worley R, Jorgensen C, Eichwald EJ (1985) Limitations of a histologic assay for syn-, allo-, and xenogeneic H-Y antigen. J Reprod Immunol 8:121–128

Download references

Author information

Correspondence to Andreas Braun.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Braun, A., Cleve, H. Unsuitability of the assay for cell-mediated lympholysis in inbred mice for H-Y antigen determination of human cells. Hum Genet 76, 369–374 (1987).

Download citation


  • Cell Proliferation
  • Internal Medicine
  • Metabolic Disease
  • Human Cell
  • Thymidine