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. 1990 Nov;9(11):3489–3496. doi: 10.1002/j.1460-2075.1990.tb07557.x

Mouse B lymphocyte specific endocytosis and recycling of MHC class II molecules.

J Salamero 1, M Humbert 1, P Cosson 1, J Davoust 1
PMCID: PMC552097  PMID: 1976509

Abstract

In B lymphocytes, the processing of exogenous proteins and the subsequent binding of antigenic peptides to class II molecules encoded by the major histocompatibility complex (MHC) occurs most likely within endocytic compartments. To examine the endocytic transport of MHC class II molecules, we used (i) surface iodination followed by internalization, pronase treatment and immunoprecipitation, (ii) in situ iodination of endosomal compartments, and (iii) confocal microscopy to visualize the fate of fluorescence coupled Fab fragments. In murine I-Ak, I-Ek positive B lymphoma cells, cell surface MHC class II molecules are partially protected from pronase digestion after 15 min at 37 degrees C and recycled back to the cell surface within the next 30 min. The fluorescence coupled Fab fragments are delivered to juxtanuclear endocytic compartments in 15 min. In contrast to the murine B cells, L fibroblasts transfected with either I-A alpha beta k or I-E alpha k beta k,d fail to internalize their surface class II molecules. A fraction of class II molecules, however, is still present in endosomal compartments as detected after in situ iodination in L fibroblasts. We conclude that the recipient L fibroblasts lack one or several factors needed for the transport of MHC class II molecules from the cell surface to the endosomes. We suggest that in murine B lymphoma cells, antigenic peptides can gain access to a pool of recycling class II molecules whereas in L cells they meet newly synthesized class II molecules targeted to the endosomal compartments.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abbas A. K., Haber S., Rock K. L. Antigen presentation by hapten-specific B lymphocytes. II. Specificity and properties of antigen-presenting B lymphocytes, and function of immunoglobulin receptors. J Immunol. 1985 Sep;135(3):1661–1667. [PubMed] [Google Scholar]
  2. Adorini L., Appella E., Doria G., Cardinaux F., Nagy Z. A. Competition for antigen presentation in living cells involves exchange of peptides bound by class II MHC molecules. Nature. 1989 Dec 14;342(6251):800–803. doi: 10.1038/342800a0. [DOI] [PubMed] [Google Scholar]
  3. Allen P. M., Unanue E. R. Differential requirements for antigen processing by macrophages for lysozyme-specific T cell hybridomas. J Immunol. 1984 Mar;132(3):1077–1079. [PubMed] [Google Scholar]
  4. Austin P., Trowsdale J., Rudd C., Bodmer W., Feldmann M., Lamb J. Functional expression of HLA-DP genes transfected into mouse fibroblasts. Nature. 1985 Jan 3;313(5997):61–64. doi: 10.1038/313061a0. [DOI] [PubMed] [Google Scholar]
  5. Blum J. S., Cresswell P. Role for intracellular proteases in the processing and transport of class II HLA antigens. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3975–3979. doi: 10.1073/pnas.85.11.3975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buus S., Werdelin O. A group-specific inhibitor of lysosomal cysteine proteinases selectively inhibits both proteolytic degradation and presentation of the antigen dinitrophenyl-poly-L-lysine by guinea pig accessory cells to T cells. J Immunol. 1986 Jan;136(2):452–458. [PubMed] [Google Scholar]
  7. Chesnut R. W., Colon S. M., Grey H. M. Requirements for the processing of antigens by antigen-presenting B cells. I. Functional comparison of B cell tumors and macrophages. J Immunol. 1982 Dec;129(6):2382–2388. [PubMed] [Google Scholar]
  8. Cresswell P. Intracellular class II HLA antigens are accessible to transferrin-neuraminidase conjugates internalized by receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8188–8192. doi: 10.1073/pnas.82.23.8188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Davidson H. W., Watts C. Epitope-directed processing of specific antigen by B lymphocytes. J Cell Biol. 1989 Jul;109(1):85–92. doi: 10.1083/jcb.109.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davis J. E., Cresswell P. Lack of detectable endocytosis of B lymphocyte MHC class II antigens using an antibody-independent technique. J Immunol. 1990 Feb 1;144(3):990–997. [PubMed] [Google Scholar]
  11. Germain R. N., Norcross M. A., Margulies D. H. Functional expression of a transfected murine class II MHC gene. Nature. 1983 Nov 10;306(5939):190–194. doi: 10.1038/306190a0. [DOI] [PubMed] [Google Scholar]
  12. Griffiths G., Hoflack B., Simons K., Mellman I., Kornfeld S. The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell. 1988 Feb 12;52(3):329–341. doi: 10.1016/s0092-8674(88)80026-6. [DOI] [PubMed] [Google Scholar]
  13. Griffiths G., Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. doi: 10.1126/science.2945253. [DOI] [PubMed] [Google Scholar]
  14. Guagliardi L. E., Koppelman B., Blum J. S., Marks M. S., Cresswell P., Brodsky F. M. Co-localization of molecules involved in antigen processing and presentation in an early endocytic compartment. Nature. 1990 Jan 11;343(6254):133–139. doi: 10.1038/343133a0. [DOI] [PubMed] [Google Scholar]
  15. Hanau D., Fabre M., Schmitt D. A., Garaud J. C., Pauly G., Tongio M. M., Mayer S., Cazenave J. P. Human epidermal Langerhans cells cointernalize by receptor-mediated endocytosis "nonclassical" major histocompatibility complex class I molecules (T6 antigens) and class II molecules (HLA-DR antigens). Proc Natl Acad Sci U S A. 1987 May;84(9):2901–2905. doi: 10.1073/pnas.84.9.2901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harding C. V., Roof R. W., Unanue E. R. Turnover of Ia-peptide complexes is facilitated in viable antigen-presenting cells: biosynthetic turnover of Ia vs. peptide exchange. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4230–4234. doi: 10.1073/pnas.86.11.4230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Harding C. V., Unanue E. R. Antigen processing and intracellular Ia. Possible roles of endocytosis and protein synthesis in Ia function. J Immunol. 1989 Jan 1;142(1):12–19. [PubMed] [Google Scholar]
  18. Haughton G., Arnold L. W., Bishop G. A., Mercolino T. J. The CH series of murine B cell lymphomas: neoplastic analogues of Ly-1+ normal B cells. Immunol Rev. 1986 Oct;93:35–51. doi: 10.1111/j.1600-065x.1986.tb01501.x. [DOI] [PubMed] [Google Scholar]
  19. Hubbard A. L., Cohn Z. A. The enzymatic iodination of the red cell membrane. J Cell Biol. 1972 Nov;55(2):390–405. doi: 10.1083/jcb.55.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knight S. C., Fryer P., Griffiths S., Harding B. Class II histocompatibility antigens on human dendritic cells. Immunology. 1987 May;61(1):21–27. [PMC free article] [PubMed] [Google Scholar]
  21. Koch N., Lipp J., Pessara U., Schenck K., Wraight C., Dobberstein B. MHC class II invariant chains in antigen processing and presentation. Trends Biochem Sci. 1989 Sep;14(9):383–386. doi: 10.1016/0968-0004(89)90013-3. [DOI] [PubMed] [Google Scholar]
  22. Kovac Z., Schwartz R. H. The molecular basis of the requirement for antigen processing of pigeon cytochrome c prior to T cell activation. J Immunol. 1985 May;134(5):3233–3240. [PubMed] [Google Scholar]
  23. Lanzavecchia A. Antigen uptake and accumulation in antigen-specific B cells. Immunol Rev. 1987 Oct;99:39–51. doi: 10.1111/j.1600-065x.1987.tb01171.x. [DOI] [PubMed] [Google Scholar]
  24. Long E. O. Intracellular traffic and antigen processing. Immunol Today. 1989 Jul;10(7):232–234. doi: 10.1016/0167-5699(89)90259-4. [DOI] [PubMed] [Google Scholar]
  25. Machy P., Barbet J., Leserman L. D. Differential endocytosis of T and B lymphocyte surface molecules evaluated with antibody-bearing fluorescent liposomes containing methotrexate. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4148–4152. doi: 10.1073/pnas.79.13.4148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Malissen B., Price M. P., Goverman J. M., McMillan M., White J., Kappler J., Marrack P., Pierres A., Pierres M., Hood L. Gene transfer of H-2 class II genes: antigen presentation by mouse fibroblast and hamster B-cell lines. Cell. 1984 Feb;36(2):319–327. doi: 10.1016/0092-8674(84)90225-3. [DOI] [PubMed] [Google Scholar]
  27. McCoy K. L., Miller J., Jenkins M., Ronchese F., Germain R. N., Schwartz R. H. Diminished antigen processing by endosomal acidification mutant antigen-presenting cells. J Immunol. 1989 Jul 1;143(1):29–38. [PubMed] [Google Scholar]
  28. Mellman I. S., Steinman R. M., Unkeless J. C., Cohn Z. A. Selective iodination and polypeptide composition of pinocytic vesicles. J Cell Biol. 1980 Sep;86(3):712–722. doi: 10.1083/jcb.86.3.712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mellman I., Fuchs R., Helenius A. Acidification of the endocytic and exocytic pathways. Annu Rev Biochem. 1986;55:663–700. doi: 10.1146/annurev.bi.55.070186.003311. [DOI] [PubMed] [Google Scholar]
  30. Neefjes J. J., Stollorz V., Peters P. J., Geuze H. J., Ploegh H. L. The biosynthetic pathway of MHC class II but not class I molecules intersects the endocytic route. Cell. 1990 Apr 6;61(1):171–183. doi: 10.1016/0092-8674(90)90224-3. [DOI] [PubMed] [Google Scholar]
  31. Nguyen Q. V., Knapp W., Humphreys R. E. Inhibition by leupeptin and antipain of the intracellular proteolysis of Ii. Hum Immunol. 1989 Mar;24(3):153–163. doi: 10.1016/0198-8859(89)90056-6. [DOI] [PubMed] [Google Scholar]
  32. Norcross M. A., Bentley D. M., Margulies D. H., Germain R. N. Membrane Ia expression and antigen-presenting accessory cell function of L cells transfected with class II major histocompatibility complex genes. J Exp Med. 1984 Nov 1;160(5):1316–1337. doi: 10.1084/jem.160.5.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nowell J., Quaranta V. Chloroquine affects biosynthesis of Ia molecules by inhibiting dissociation of invariant (gamma) chains from alpha-beta dimers in B cells. J Exp Med. 1985 Oct 1;162(4):1371–1376. doi: 10.1084/jem.162.4.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ozato K., Mayer N., Sachs D. H. Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. J Immunol. 1980 Feb;124(2):533–540. [PubMed] [Google Scholar]
  35. Pierres M., Devaux C., Dosseto M., Marchetto S. Clonal analysis of B- and T-cell responses to Ia antigens. I. Topology of epitope regions on I-Ak and I-Ek molecules analyzed with 35 monoclonal alloantibodies. Immunogenetics. 1981 Dec;14(6):481–495. doi: 10.1007/BF00350120. [DOI] [PubMed] [Google Scholar]
  36. Pollerberg G. E., Schachner M., Davoust J. Differentiation state-dependent surface mobilities of two forms of the neural cell adhesion molecule. Nature. 1986 Dec 4;324(6096):462–465. doi: 10.1038/324462a0. [DOI] [PubMed] [Google Scholar]
  37. Roche P. A., Cresswell P. Invariant chain association with HLA-DR molecules inhibits immunogenic peptide binding. Nature. 1990 Jun 14;345(6276):615–618. doi: 10.1038/345615a0. [DOI] [PubMed] [Google Scholar]
  38. Rock K. L., Benacerraf B., Abbas A. K. Antigen presentation by hapten-specific B lymphocytes. I. Role of surface immunoglobulin receptors. J Exp Med. 1984 Oct 1;160(4):1102–1113. doi: 10.1084/jem.160.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Roederer M., Bowser R., Murphy R. F. Kinetics and temperature dependence of exposure of endocytosed material to proteolytic enzymes and low pH: evidence for a maturation model for the formation of lysosomes. J Cell Physiol. 1987 May;131(2):200–209. doi: 10.1002/jcp.1041310209. [DOI] [PubMed] [Google Scholar]
  40. Roosnek E., Demotz S., Corradin G., Lanzavecchia A. Kinetics of MHC-antigen complex formation on antigen-presenting cells. J Immunol. 1988 Jun 15;140(12):4079–4082. [PubMed] [Google Scholar]
  41. Sant A. J., Cullen S. E., Schwartz B. D. Biosynthetic relationships of the chondroitin sulfate proteoglycan with Ia and invariant chain glycoproteins. J Immunol. 1985 Jul;135(1):416–422. [PubMed] [Google Scholar]
  42. Schmid S. L., Fuchs R., Male P., Mellman I. Two distinct subpopulations of endosomes involved in membrane recycling and transport to lysosomes. Cell. 1988 Jan 15;52(1):73–83. doi: 10.1016/0092-8674(88)90532-6. [DOI] [PubMed] [Google Scholar]
  43. Seglen P. O. Inhibitors of lysosomal function. Methods Enzymol. 1983;96:737–764. doi: 10.1016/s0076-6879(83)96063-9. [DOI] [PubMed] [Google Scholar]
  44. Shastri N., Malissen B., Hood L. Ia-transfected L-cell fibroblasts present a lysozyme peptide but not the native protein to lysozyme-specific T cells. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5885–5889. doi: 10.1073/pnas.82.17.5885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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. doi: 10.1084/jem.158.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Simonis S., Miller J., Cullen S. E. The role of the Ia-invariant chain complex in the posttranslational processing and transport of Ia and invariant chain glycoproteins. J Immunol. 1989 Dec 1;143(11):3619–3625. [PubMed] [Google Scholar]
  47. Stockinger B., Pessara U., Lin R. H., Habicht J., Grez M., Koch N. A role of Ia-associated invariant chains in antigen processing and presentation. Cell. 1989 Feb 24;56(4):683–689. doi: 10.1016/0092-8674(89)90590-4. [DOI] [PubMed] [Google Scholar]
  48. Streicher H. Z., Berkower I. J., Busch M., Gurd F. R., Berzofsky J. A. Antigen conformation determines processing requirements for T-cell activation. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6831–6835. doi: 10.1073/pnas.81.21.6831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Suissa M. Spectrophotometric quantitation of silver grains eluted from autoradiograms. Anal Biochem. 1983 Sep;133(2):511–514. doi: 10.1016/0003-2697(83)90117-3. [DOI] [PubMed] [Google Scholar]
  50. Townsend A., Ohlén C., Bastin J., Ljunggren H. G., Foster L., Kärre K. Association of class I major histocompatibility heavy and light chains induced by viral peptides. Nature. 1989 Aug 10;340(6233):443–448. doi: 10.1038/340443a0. [DOI] [PubMed] [Google Scholar]
  51. Ziegler H. K., Unanue E. R. 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. doi: 10.1073/pnas.79.1.175. [DOI] [PMC free article] [PubMed] [Google Scholar]

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