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The requirements for antigen multivalency in class I antigen recognition and triggering of primed precursor cytolytic T lymphocytes.

S H Herrmann and M F Mescher
J Immunol April 15, 1986, 136 (8) 2816-2825;
S H Herrmann
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M F Mescher
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Abstract

In vitro generation of a secondary cytolytic T lymphocyte (CTL) response to Class I alloantigen requires two signals: recognition of the Class I antigen by precursor CTL (Signal 1), and subsequent interaction with lymphokine(s) (Signal 2). Previous work using subcellular antigen stimulation has demonstrated that the required lymphokine(s) is produced as a result of adherent cell uptake, processing, and Ia-restricted presentation of alloantigen to helper T cells. This pathway could be bypassed by addition to the cultures of supernatant from Con A-stimulated rat spleen cells. When an optimal level of lymphokine(s) is provided by addition of Con A supernatant, the magnitude of the CTL response obtained is dependent on the effectiveness of alloantigen recognition and triggering of the primed precursor CTL (pCTL). By using this approach, we examined the cellular and molecular requirements for generation of Signal 1. Previous results had indicated that pCTL were able to directly recognize subcellular antigen, and that cellular presentation of the antigen to pCTL was not required. Further evidence for this was provided by the finding that pulsing of the responder population for short times with liposomes containing purified H-2Kk resulted in effective stimulation of the response. Exposure of cells to antigen for 1 to 2 hr at 4 degrees C generated responses of comparable magnitude to those obtained when antigen was continuously present in the cultures. Experiments were also done to directly examine the ability of alloantigen-pulsed splenic adherent cells (SAC) to deliver Signal 1. Although the antigen-pulsed SAC were very effective in presenting to helper T cells to result in factor production, they were found to be very ineffective in providing Signal 1 to the pCTL. Having obtained strong evidence for triggering of pCTL occurring via direct recognition of the subcellular alloantigen, we then examined the role of antigen multivalency in recognition and triggering. Purified H-2Kk was prepared in a variety of forms of differing multivalency, ranging from monovalent papain cleavage product to large, highly multivalent liposomes and plasma membranes. The magnitude of the CTL responses obtained was found to be critically dependent on the multivalency of the antigen preparation. Examination of the antigen dose-response curves and maximal responses obtained suggests that valency of the antigen may be important both in determining the avidity of interaction between the pCTL and the antigen-bearing structure, and in determining the extent to which localized receptor cross-linking occurs on the cell surface to result in triggering.

  • Copyright © 1986 by American Association of Immunologists

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The Journal of Immunology
Vol. 136, Issue 8
15 Apr 1986
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The requirements for antigen multivalency in class I antigen recognition and triggering of primed precursor cytolytic T lymphocytes.
S H Herrmann, M F Mescher
The Journal of Immunology April 15, 1986, 136 (8) 2816-2825;

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The requirements for antigen multivalency in class I antigen recognition and triggering of primed precursor cytolytic T lymphocytes.
S H Herrmann, M F Mescher
The Journal of Immunology April 15, 1986, 136 (8) 2816-2825;
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Print ISSN 0022-1767        Online ISSN 1550-6606