This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Rajagopalan, G. Articles by David, C. S. Search for Related Content PubMed PubMed Citation Articles by Rajagopalan, G. Articles by David, C. S.

Endogenous Superantigens Shape Response to Exogenous Superantigens

Department of Immunology,¹ Division of Nephrology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, Minnesota²

Received 23 February 2005/ Returned for modification 26 April 2005/ Accepted 28 June 2005

	ABSTRACT

Top Abstract Text References

 
Endogenous superantigen-mediated thymic negative selection resulted in a paucity of mature T cells bearing T-cell receptor (TCR) Vß8 in the periphery. Consequently, the magnitude of immune response to exogenous superantigen staphylococcal enterotoxin B, which activates TCR Vß8⁺ T cells, was significantly reduced and conferred protection from superantigen-induced mortality.

	TEXT

Top Abstract Text References

 
Superantigens are a family of microbial proteins that bind directly to major histocompatibility complex (MHC) class II molecules outside of the peptide-binding groove and activate CD4⁺ and CD8⁺ T cells expressing certain T-cell receptor (TCR) Vß families (10). Superantigens encoded by retroviruses integrated into the genome are called "endogenous superantigens," and those of bacterial origin are called "exogenous superantigens" (3). The outcome of superantigen-mediated activation differs for mature T cells and immature thymocytes. While the former proliferate vigorously, the latter rapidly undergo apoptosis. Therefore, thymic expression of endogenous superantigens encoded by integrated retrovirus transmitted through the germ line causes deletion of thymocytes bearing certain TCR Vß families, resulting in a paucity of those TCR Vß families in the mature peripheral T-cell pool (13).

Endogenous superantigen-mediated TCR Vß deletions are reported in humans (8). HERV-K18, a human endogenous retrovirus-encoded superantigen, can cause deletion of TCR Vß7⁺ thymocytes (7). Incidentally, TCR Vß7-bearing T cells are strongly stimulated by the exogenous superantigen staphylococcal enterotoxin A (SEA) in humans (10). Individuals expressing HERV-K18 in the thymus would theoretically mount a poor immune response to SEA due to a dearth of mature TCR Vß7⁺ T cells in the periphery and thus could be protected from SEA-induced toxic shock syndrome. Due to the extreme toxicity of bacterial superantigens to humans, we tested the hypothesis that endogenous superantigen-mediated TCR Vß deletions can modulate the immune response elicited by an exogenous bacterial superantigen using the well-established HLA class II transgenic mouse model (2, 11, 14, 15).

We have generated two independent lines of HLA class II transgenic mice expressing HLA-DR3 (DRB1*0301) (12) and HLA-DR2 (DRB1*1501) (1). These two HLA-DR alleles share the same DR chain, DRA*0103. Previous studies have shown that expression of transgenic class II molecules in these mice is comparable with murine MHC class II molecules. However, it should be noted that these mice completely lack all endogenous MHC class II genes (6), and the T-cell responses are restricted only by the transgenic HLA class II (1). While the levels of expression of transgenic class II (Fig. 1A) and distribution of CD4⁺ and CD8⁺ T cells (Fig. 1B) are comparable between these lines, the presence of some endogenous superantigen in the DR2 lines deleted CD4⁺ as well as CD8⁺ T cells bearing TCR Vß8 in thymus, resulting in negligible representation of this TCR in the periphery (Fig. 2). Deletion of TCR Vß8 in thymus is dependent on the expression of a functional class II molecule, as mice expressing the DR chain alone without the HLA-DRB1*1501 ß chain do not delete TCR Vß8 (Fig. 3). Since this deletion is MHC class II dependent and occurs in both CD4⁺ and CD8⁺ T cells during transition from the double-positive (DP) to the single-positive stage in thymus, we conclude that this deletion is mediated by endogenous superantigen.

View larger version (14K): [in this window] [in a new window]   FIG. 1. MHC class II expression and T-cell development in HLA-DR transgenic mice. (A) Splenocytes from age-matched HLA-DR3 (DRB*0301) and HLA-DR2 (DRB*1501) transgenic mice (n 4 mice/group) expressing common DRA*0103 were stained with L227 (anti-DR antibody) to check expression of transgenic HLA-DR by flow cytometry. Shown are percentages of cells expressing DR. (B) CD4+ and CD8+ T-cell development in DR3 and DR2 mice (n 4 mice/group) as determined by flow cytometry. Shown are percentages of cells positive for CD4 or CD8.

View larger version (17K): [in this window] [in a new window]   FIG. 2. Thymic selection in HLA-DR transgenic mice. Thymocytes from age-matched HLA-DR3 mice (n 4 mice/group) expressing both DR and DRß chains and HLA-DR2 transgenic mice expressing the DR chain but expressing or lacking the DRß chain were analyzed by flow cytometry for CD4 and CD8 coreceptors. Note the significant presence of TCR Vß8+ T cells among the single- positive (SP) subsets in DRß chain-negative DR2 littermates and the absence of TCR Vß8+ T cells in DRß chain-positive DR2 littermates, indicating the requirement of functional HLA class II molecules for thymic negative selection. Only CD8 SP cells are analyzed, as CD4 SP cells would be absent in the absence of functional class II molecules.

View larger version (15K): [in this window] [in a new window]   FIG. 3. TCR Vß repertoire in HLA-DR transgenic mice. Splenocytes from age-matched HLA-DR3 and -DR2 transgenic mice (n = 3 to 6 mice/group) were stained for CD4, CD8, and TCR Vß-specific fluorochrome-conjugated antibodies and analyzed by flow cytometry. Shown are percentages of cells expressing a specific TCR Vß family within the respective gated population.

Contrary to the expectation that the absence of TCR Vß8-bearing T cells in HLA-DR2 mice would result in substantial reduction in SEB-induced immune activation, we observed that splenocytes from HLA-DR2 mice showed a strong in vitro T-cell response to SEB, albeit lesser than DR3 mice (Fig. 4). To gain a better insight into this process, HLA-DR3 and HLA-DR2 mice were challenged in vivo with purified SEB and the TCR Vß usage was determined by flow cytometry 3 days later. In HLA-DR3 transgenic mice, as shown by us earlier, TCR Vß8⁺ T cells increased in both CD4⁺ and CD8⁺ subsets (Fig. 5). However, in HLA-DR2 transgenic mice, since TCR Vß8-bearing T cells are deleted, they did not contribute much to the peripheral T-cell pool. However, T cells bearing TCR Vß7 expanded by nearly 10-fold in both CD8⁺ and CD4⁺ T-cell populations. In addition, T cells bearing TCR Vß14, which did not respond in DR3 mice, expanded significantly in response to SEB in DR2 mice. Together, TCR Vß7 and Vß14 contributed to 55 to 60% of the T-cell pool following SEB injection, whereas they contributed to less than 25% of the CD4⁺ and CD8⁺ T-cell pool in naive mice. Similar results were obtained with another DR2 transgenic line expressing the same DR chain but a different DRß chain (DRB*1502), which also shows TCR Vß8 family deletion (data not shown).

View larger version (11K): [in this window] [in a new window]   FIG. 4. In vitro T-cell response to SEB. Splenocytes from age-matched HLA-DR3 and -DR2 transgenic mice (n = 3 to 6 mice/group) were cultured in vitro with the indicated concentrations of SEB for 42 h. Cell proliferation was determined by thymidine incorporation assay.

View larger version (17K): [in this window] [in a new window]   FIG. 5. TCR Vß repertoire in superantigen-challenged HLA-DR transgenic mice. Splenocytes from age-matched HLA-DR3 and -DR2 transgenic mice (n = 3 to 10 mice/group) challenged with 10 µg of SEB 3 days earlier were stained for CD4, CD8, and TCR Vß-specific fluorochrome-conjugated antibodies and analyzed by flow cytometry. Shown are percentages of cells expressing specific a TCR Vß family within the respective gated population.

View larger version (13K): [in this window] [in a new window]   FIG. 6. SEB-induced thymic deletion in HLA-DR transgenic mice. Thymocytes from age-matched HLA-DR3 and -DR2 transgenic mice (n = 3 to 10 mice/group) challenged with 10 µg of SEB 3 days earlier were stained for CD4 as well as CD8 and analyzed by flow cytometry. Shown are the reductions in single-positive (SP) and DP thymocyte subsets.

	ACKNOWLEDGMENTS

 
This study was supported by NIH grant AI14764 to C.S.D. G.R. is a recipient of a Juvenile Diabetes Research Foundation fellowship.

We thank Julie Hanson and her crew for excellent mouse husbandry and Michele Smart for typing the mice.

	FOOTNOTES

 
* Corresponding author. Mailing address: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, 200 First Street, SW, Rochester, MN 55905. Phone: (507) 284-8180. Fax: (507) 266-0981. E-mail: david.chella{at}mayo.edu.

	REFERENCES

Top Abstract Text References

 

1. Cheng, S., M. Smart, J. Hanson, and C. S. David. 2003. Characterization of HLA DR2 and DQ8 transgenic mouse with a new engineered mouse class II deletion, which lacks all endogenous class II genes. J. Autoimmun. 21:195-199.[CrossRef][Medline]
2. DaSilva, L., B. C. Welcher, R. G. Ulrich, M. J. Aman, C. S. David, and S. Bavari. 2002. Humanlike immune response of human leukocyte antigen-DR3 transgenic mice to staphylococcal enterotoxins: a novel model for superantigen vaccines. J. Infect. Dis. 185:1754-1760.[CrossRef][Medline]
3. Herman, A., J. W. Kappler, P. Marrack, and A. M. Pullen. 1991. Superantigens: mechanism of T-cell stimulation and role in immune responses. Annu. Rev. Immunol. 9:745-772.[Medline]
4. Kishimoto, H., C. D. Surh, and J. Sprent. 1998. A role for Fas in negative selection of thymocytes in vivo. J. Exp. Med. 187:1427-1438.[Abstract/Free Full Text]
5. Li, H., A. Llera, E. L. Malchiodi, and R. A. Mariuzza. 1999. The structural basis of T cell activation by superantigens. Annu. Rev. Immunol. 17:435-466.[CrossRef][Medline]
6. Madsen, L., N. Labrecque, J. Engberg, A. Dierich, A. Svejgaard, C. Benoist, D. Mathis, and L. Fugger. 1999. Mice lacking all conventional MHC class II genes. Proc. Natl. Acad. Sci. USA 96:10338-10343.[Abstract/Free Full Text]
7. Meylan, F., M. De Smedt, G. Leclercq, J. Plum, O. Leupin, S. Marguerat, and B. Conrad. 2005. Negative thymocyte selection to HERV-K18 superantigens in humans. Blood 105:4377-4382.[Abstract/Free Full Text]
8. Nelson, P. N., P. Hooley, D. Roden, H. Davari Ejtehadi, P. Rylance, P. Warren, J. Martin, and P. G. Murray. 2004. Human endogenous retroviruses: transposable elements with potential? Clin. Exp. Immunol. 138:1-9.[CrossRef][Medline]
9. Portis, J. L. 2002. Perspectives on the role of endogenous human retroviruses in autoimmune diseases. Virology 296:1-5.[CrossRef][Medline]
10. Proft, T., and J. D. Fraser. 2003. Bacterial superantigens. Clin. Exp. Immunol. 133:299-306.[CrossRef][Medline]
11. Rajagopalan, G., M. Sen, and C. S. David. 2004. In vitro and in vivo evaluation of staphylococcal superantigen peptide antagonisits. Infect. Immun. 72:6733-6737.[Abstract/Free Full Text]
12. Rajagopalan, G., M. K. Smart, S. Cheng, C. J. Krco, K. L. Johnson, and C. S. David. 2003. Expression and function of HLA-DR3 and DQ8 in transgenic mice lacking functional H2-M. Tissue Antigens 62:149-161.[CrossRef][Medline]
13. Simpson, E., K. Takacs, and D. M. Altmann. 1994. Thymic repertoire selection by superantigens: presentation by human and mouse MHC molecules. Thymus 23:1-13.[Medline]
14. Sriskandan, S., M. Unnikrishnan, T. Krausz, H. Dewchand, S. Van Noorden, J. Cohen, and D. M. Altmann. 2001. Enhanced susceptibility to superantigen-associated streptococcal sepsis in human leukocyte antigen-DQ transgenic mice. J. Infect. Dis. 184:166-173.[CrossRef][Medline]
15. Yeung, R. S., J. M. Penninger, T. Kundig, W. Khoo, P. S. Ohashi, G. Kroemer, and T. W. Mak. 1996. Human CD4 and human major histocompatibility complex class II (DQ6) transgenic mice: supersensitivity to superantigen-induced septic shock. Eur. J. Immunol. 26:1074-1082.[Medline]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Rajagopalan, G. Articles by David, C. S. Search for Related Content PubMed PubMed Citation Articles by Rajagopalan, G. Articles by David, C. S.