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J Exp Med. 1995 January 1; 181(1): 223–234.
PMCID: PMC2191808

The requirement for DM in class II-restricted antigen presentation and SDS-stable dimer formation is allele and species dependent

Abstract

Recently several cell lines have been identified with mutations in a major histocompatibility complex (MHC)-linked protein that lead to defects in class II-restricted antigen presentation and a defect in the formation of class II SDS-stable dimers. The defect in these cells has recently been shown to result from the inability to express the MHC- encoded nonclassical class II molecule called DM. To further examine the role of DM in class II-restricted antigen presentation, we asked if this defect would equally affect different allelic and species variants of class II molecules. To investigate this, we transfected the parent cell lines T1 and 8.1.6 and their respective antigen presentation mutants T2 and 9.5.3 with the genes encoding I-Ad and examined the derived transfectants for their ability to present antigen, the conformation of I-Ad at the cell surface, association of I-Ad with invariant chain (Ii), and the ability to form I-Ad SDS-stable dimers. The lack of functional DM expression did not affect any of the anti-I- Ad monoclonal antibody (mAb) epitopes tested or the ability of I-Ad to associate and dissociate with Ii. Surprisingly, these studies also revealed that the antigen presentation defect observed for DR in the 9.5.3 cells did not compromise I-Ad-restricted antigen presentation. In addition, we found that the level of SDS-stable dimer formation did not correlate with antigen presentation capacity for I-Ad and that the amount of SDS-stable I-Ad dimer depends on the cellular context in which the class II molecule is expressed. Our results suggest that the ability to form SDS-stable dimer is not strictly correlated with class II-restricted antigen presentation. Finally, when two allelic forms of murine class II molecules were compared in the defective T2 cell line, it was found that I-Ak but not I-Ad forms SDS-stable dimers equivalent to that seen in the parental cell lines. Overall, our results suggest that DM may modulate rather than play a requisite role in I-Ad- restricted antigen presentation and SDS-stable dimer formation and that dependency on DM may be allele or species specific.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ziegler HK, Unanue ER. Decrease in macrophage antigen catabolism caused by ammonia and chloroquine is associated with inhibition of antigen presentation to T cells. Proc Natl Acad Sci U S A. 1982 Jan;79(1):175–178. [PubMed]
  • Peters PJ, Neefjes JJ, Oorschot V, Ploegh HL, Geuze HJ. Segregation of MHC class II molecules from MHC class I molecules in the Golgi complex for transport to lysosomal compartments. Nature. 1991 Feb 21;349(6311):669–676. [PubMed]
  • Tulp A, Verwoerd D, Dobberstein B, Ploegh HL, Pieters J. Isolation and characterization of the intracellular MHC class II compartment. Nature. 1994 May 12;369(6476):120–126. [PubMed]
  • Amigorena S, Drake JR, Webster P, Mellman I. Transient accumulation of new class II MHC molecules in a novel endocytic compartment in B lymphocytes. Nature. 1994 May 12;369(6476):113–120. [PubMed]
  • West MA, Lucocq JM, Watts C. Antigen processing and class II MHC peptide-loading compartments in human B-lymphoblastoid cells. Nature. 1994 May 12;369(6476):147–151. [PubMed]
  • Qiu Y, Xu X, Wandinger-Ness A, Dalke DP, Pierce SK. Separation of subcellular compartments containing distinct functional forms of MHC class II. J Cell Biol. 1994 May;125(3):595–605. [PMC free article] [PubMed]
  • Sant AJ, Miller J. MHC class II antigen processing: biology of invariant chain. Curr Opin Immunol. 1994 Feb;6(1):57–63. [PubMed]
  • Sadegh-Nasseri S, Germain RN. A role for peptide in determining MHC class II structure. Nature. 1991 Sep 12;353(6340):167–170. [PubMed]
  • Germain RN, Hendrix LR. MHC class II structure, occupancy and surface expression determined by post-endoplasmic reticulum antigen binding. Nature. 1991 Sep 12;353(6340):134–139. [PubMed]
  • Srinivasan M, Marsh EW, Pierce SK. Characterization of naturally processed antigen bound to major histocompatibility complex class II molecules. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7928–7932. [PubMed]
  • Nelson CA, Petzold SJ, Unanue ER. Identification of two distinct properties of class II major histocompatibility complex-associated peptides. Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1227–1231. [PubMed]
  • Davidson HW, Reid PA, Lanzavecchia A, Watts C. Processed antigen binds to newly synthesized MHC class II molecules in antigen-specific B lymphocytes. Cell. 1991 Oct 4;67(1):105–116. [PubMed]
  • Roche PA, Cresswell P. Invariant chain association with HLA-DR molecules inhibits immunogenic peptide binding. Nature. 1990 Jun 14;345(6276):615–618. [PubMed]
  • Teyton L, O'Sullivan D, Dickson PW, Lotteau V, Sette A, Fink P, Peterson PA. Invariant chain distinguishes between the exogenous and endogenous antigen presentation pathways. Nature. 1990 Nov 1;348(6296):39–44. [PubMed]
  • DeMars R, Chang CC, Shaw S, Reitnauer PJ, Sondel PM. Homozygous deletions that simultaneously eliminate expressions of class I and class II antigens of EBV-transformed B-lymphoblastoid cells. I. Reduced proliferative responses of autologous and allogeneic T cells to mutant cells that have decreased expression of class II antigens. Hum Immunol. 1984 Oct;11(2):77–97. [PubMed]
  • Mellins E, Kempin S, Smith L, Monji T, Pious D. A gene required for class II-restricted antigen presentation maps to the major histocompatibility complex. J Exp Med. 1991 Dec 1;174(6):1607–1615. [PMC free article] [PubMed]
  • Cotner T, Mellins E, Johnson AH, Pious D. Mutations affecting antigen processing impair class II-restricted allorecognition. J Immunol. 1991 Jan 15;146(2):414–417. [PubMed]
  • Riberdy JM, Cresswell P. The antigen-processing mutant T2 suggests a role for MHC-linked genes in class II antigen presentation. J Immunol. 1992 Apr 15;148(8):2586–2590. [PubMed]
  • Riberdy JM, Newcomb JR, Surman MJ, Barbosa JA, Cresswell P. HLA-DR molecules from an antigen-processing mutant cell line are associated with invariant chain peptides. Nature. 1992 Dec 3;360(6403):474–477. [PubMed]
  • Ceman S, Petersen JW, Pinet V, DeMars R. Gene required for normal MHC class II expression and function is localized to approximately 45 kb of DNA in the class II region of the MHC. J Immunol. 1994 Mar 15;152(6):2865–2873. [PubMed]
  • Morris P, Shaman J, Attaya M, Amaya M, Goodman S, Bergman C, Monaco JJ, Mellins E. An essential role for HLA-DM in antigen presentation by class II major histocompatibility molecules. Nature. 1994 Apr 7;368(6471):551–554. [PubMed]
  • Fling SP, Arp B, Pious D. HLA-DMA and -DMB genes are both required for MHC class II/peptide complex formation in antigen-presenting cells. Nature. 1994 Apr 7;368(6471):554–558. [PubMed]
  • Johnson JP, Meo T, Riethmüller G, Schendel DJ, Wank R. Direct demonstration of an HLA-DR allotypic determinant on the low molecular weight (beta) subunit using a mouse monoclonal antibody specific for DR3. J Exp Med. 1982 Jul 1;156(1):104–111. [PMC free article] [PubMed]
  • Loss GE, Jr, Elias CG, Fields PE, Ribaudo RK, McKisic M, Sant AJ. Major histocompatibility complex class II-restricted presentation of an internally synthesized antigen displays cell-type variability and segregates from the exogenous class II and endogenous class I presentation pathways. J Exp Med. 1993 Jul 1;178(1):73–85. [PMC free article] [PubMed]
  • Gunning P, Leavitt J, Muscat G, Ng SY, Kedes L. A human beta-actin expression vector system directs high-level accumulation of antisense transcripts. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4831–4835. [PubMed]
  • Sant AJ, Germain RN. Intracellular competition for component chains determines class II MHC cell surface phenotype. Cell. 1989 Jun 2;57(5):797–805. [PubMed]
  • Shimonkevitz R, Kappler J, Marrack P, Grey H. Antigen recognition by H-2-restricted T cells. I. Cell-free antigen processing. J Exp Med. 1983 Aug 1;158(2):303–316. [PMC free article] [PubMed]
  • Braunstein NS, Germain RN, Loney K, Berkowitz N. Structurally interdependent and independent regions of allelic polymorphism in class II MHC molecules. Implications for Ia function and evolution. J Immunol. 1990 Sep 15;145(6):1635–1645. [PubMed]
  • Steinman RM, Nogueira N, Witmer MD, Tydings JD, Mellman IS. Lymphokine enhances the expression and synthesis of Ia antigens on cultured mouse peritoneal macrophages. J Exp Med. 1980 Nov 1;152(5):1248–1261. [PMC free article] [PubMed]
  • Lampson LA, Levy R. Two populations of Ia-like molecules on a human B cell line. J Immunol. 1980 Jul;125(1):293–299. [PubMed]
  • Sarmiento M, Loken MR, Fitch FW. Structural differences in cell surface T25 polypeptides from thymocytes and cloned T cells. Hybridoma. 1981;1(1):13–26. [PubMed]
  • Ozato K, Mayer N, Sachs DH. Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. J Immunol. 1980 Feb;124(2):533–540. [PubMed]
  • Sant AJ. Isotypic residues in the membrane proximal domain of MHC class II beta-chains control activation of CD4+ T cells. J Immunol. 1993 Jun 15;150(12):5299–5310. [PubMed]
  • Peterson M, Miller J. Antigen presentation enhanced by the alternatively spliced invariant chain gene product p41. Nature. 1992 Jun 18;357(6379):596–598. [PubMed]
  • Sant AJ, Hendrix LR, Coligan JE, Maloy WL, Germain RN. Defective intracellular transport as a common mechanism limiting expression of inappropriately paired class II major histocompatibility complex alpha/beta chains. J Exp Med. 1991 Oct 1;174(4):799–808. [PMC free article] [PubMed]
  • Pinet V, Malnati MS, Long EO. Two processing pathways for the MHC class II-restricted presentation of exogenous influenza virus antigen. J Immunol. 1994 May 15;152(10):4852–4860. [PubMed]
  • Chicz RM, Urban RG, Lane WS, Gorga JC, Stern LJ, Vignali DA, Strominger JL. Predominant naturally processed peptides bound to HLA-DR1 are derived from MHC-related molecules and are heterogeneous in size. Nature. 1992 Aug 27;358(6389):764–768. [PubMed]
  • Chicz RM, Urban RG, Gorga JC, Vignali DA, Lane WS, Strominger JL. Specificity and promiscuity among naturally processed peptides bound to HLA-DR alleles. J Exp Med. 1993 Jul 1;178(1):27–47. [PMC free article] [PubMed]
  • Hunt DF, Michel H, Dickinson TA, Shabanowitz J, Cox AL, Sakaguchi K, Appella E, Grey HM, Sette A. Peptides presented to the immune system by the murine class II major histocompatibility complex molecule I-Ad. Science. 1992 Jun 26;256(5065):1817–1820. [PubMed]
  • Lanzavecchia A, Reid PA, Watts C. Irreversible association of peptides with class II MHC molecules in living cells. Nature. 1992 May 21;357(6375):249–252. [PubMed]
  • Germain RN, Rinker AG., Jr Peptide binding inhibits protein aggregation of invariant-chain free class II dimers and promotes surface expression of occupied molecules. Nature. 1993 Jun 24;363(6431):725–728. [PubMed]
  • Sette A, Ceman S, Kubo RT, Sakaguchi K, Appella E, Hunt DF, Davis TA, Michel H, Shabanowitz J, Rudersdorf R, et al. Invariant chain peptides in most HLA-DR molecules of an antigen-processing mutant. Science. 1992 Dec 11;258(5089):1801–1804. [PubMed]
  • Peterson M, Miller J. Invariant chain influences the immunological recognition of MHC class II molecules. Nature. 1990 May 10;345(6271):172–174. [PubMed]
  • Nadimi F, Moreno J, Momburg F, Heuser A, Fuchs S, Adorini L, Hämmerling GJ. Antigen presentation of hen egg-white lysozyme but not of ribonuclease A is augmented by the major histocompatibility complex class II-associated invariant chain. Eur J Immunol. 1991 May;21(5):1255–1263. [PubMed]
  • Bertolino P, Forquet F, Pont S, Koch N, Gerlier D, Rabourdin-Combe C. Correlation between invariant chain expression level and capability to present antigen to MHC class II-restricted T cells. Int Immunol. 1991 May;3(5):435–443. [PubMed]
  • Momburg F, Fuchs S, Drexler J, Busch R, Post M, Hämmerling GJ, Adorini L. Epitope-specific enhancement of antigen presentation by invariant chain. J Exp Med. 1993 Oct 1;178(4):1453–1458. [PMC free article] [PubMed]
  • Layet C, Germain RN. Invariant chain promotes egress of poorly expressed, haplotype-mismatched class II major histocompatibility complex A alpha A beta dimers from the endoplasmic reticulum/cis-Golgi compartment. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2346–2350. [PubMed]
  • Anderson MS, Miller J. Invariant chain can function as a chaperone protein for class II major histocompatibility complex molecules. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2282–2286. [PubMed]

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