8536d_ch08_185-199 8/2/02 10:08 AM Page 186 mac79 Mac 79: 45_Bwppldsby et al./ Immunology Se 186 PART 11 Generation of B-Cell and T-Cell Response y LCM virus Strain 2 or 13 Strain 2 or 13 orC2×13)F1 orC2×13)F1 Peritoneal exudate cells Lymph node cells (containing Tc cells) (retains Peritoneal macrophages H-2k target cells H-2 LCM-infected H-2b LCM-infected target cells arget cells Antigen-pulsed Antigen-primed T-cells ⊙⊙⊙ B!:8 51Cr release +51Cr release 51 Cr release (no lysis) lysis) (no lysis) FICURE8-2 Classic experiment of Zinkernagel and Doherty demonstrating that antigen recognition by Tc cells exhibits MHC re- strictionH-2 mice were primed with the lymphocytic choriomeni gitis(LCM)virus to induce cytotoxic T lymphocytes(CTLs) specific Measure t-cell for the virus Spleen cells from this LCM-primed mouse were then added to target cells of different H-2 haplotypes that were intracellu- larly labeled withCr(black dots)and either infected or not with the LCM virus CTL-mediated killing of the target cells, as measured by Antigen-primed Antigen-pulsed macrophages the release of Cr into the culture supernatant, occurred only if the Strain 2 Strain 13(2x 13)F, target cells were infected with LCM and had the same MHC haplo- ype as the CTLs. ADapted from P C. Doherty and R. M. Zinkemagel, Strain 13 1975,.Exp.Med.141:502 (2×13)F1 restricted. In 1996, Doherty and Zinkernagel were awarded FIGURE8-1Experimental demonstration of self-MHC restriction of the Nobel prize for their major contribution to the under TH cells. Peritoneal exudate cells from strain 2, strain 13, or(2 X 13)F1 standing of cell-mediated immunity. guinea pigs were incubated in plastic Petri dishes, allowing enrichment of macrophages, which are adherent cells. The peritoneal macro- phages were then incubated with antigen. These "antigen-pulse macrophages were incubated in vitro with T cells from strain 2, strain Role of Antigen-Presenting Cells 13, or(2X 13)F1 guinea pigs, and the degree of T-cell proliferation As early as 1959, immunologists were confronted with data was assessed. The results indicated that TH cells could proliferate only suggesting that T cells and B cells recognized antigen by dif- in response to antigen presented by macrophages that shared MHC al- ferent mechanisms. The dogma of the time, which persisted leles.(Adapted from A Rosenthal and E Shevach, 1974, J. Exp. Med. until the 1980s, was that cells of the immune system recog- 138: 1194, by copyright permission of the Rockefeller University Press. I nize the entire protein in its native conformation. However, experiments by P G H. Gell and B Benacerraf demonstrated that, while a primary antibody response and cell-mediated that the Tc cell and the virus-infected target cell must share response were induced by a protein in its native conforma class I molecules encoded by the K or D regions of the MHC. tion, a secondary antibody response(mediated by B cells) Thus, antigen recognition by CD8 Tc cells is class I MHc could be induced only by native antigen, whereas a secondaryrestricted. In 1996, Doherty and Zinkernagel were awarded the Nobel prize for their major contribution to the under￾standing of cell-mediated immunity. Role of Antigen-Presenting Cells As early as 1959, immunologists were confronted with data suggesting that T cells and B cells recognized antigen by dif￾ferent mechanisms. The dogma of the time, which persisted until the 1980s, was that cells of the immune system recog￾nize the entire protein in its native conformation. However, experiments by P. G. H. Gell and B. Benacerraf demonstrated that, while a primary antibody response and cell-mediated response were induced by a protein in its native conforma￾tion, a secondary antibody response (mediated by B cells) could be induced only by native antigen, whereas a secondary 186 PART II Generation of B-Cell and T-Cell Responses Antigen-pulsed macrophages Antigen-primed T cell Strain 2 Strain 13 (2 × 13)F1 Strain 2 Strain 13 (2 × 13)F1 + + − + − + + + + Strain 2 or 13 or (2 × 13)F1 Strain 2 or 13 or (2 × 13)F1 Antigen Peritoneal exudate cells Peritoneal macrophages Adherent cells Antigen Antigen-pulsed macrophages Measure T-cell proliferation Lymph node cells Antigen-primed T-cells Adherence column (retains macrophages) 7 days FIGURE 8-1 Experimental demonstration of self-MHC restriction of TH cells. Peritoneal exudate cells from strain 2, strain 13, or (2 13) F1 guinea pigs were incubated in plastic Petri dishes, allowing enrichment of macrophages, which are adherent cells. The peritoneal macro￾phages were then incubated with antigen. These “antigen-pulsed” macrophages were incubated in vitro with T cells from strain 2, strain 13, or (2 13) F1 guinea pigs, and the degree of T-cell proliferation was assessed. The results indicated that TH cells could proliferate only in response to antigen presented by macrophages that shared MHC al￾leles. [Adapted from A. Rosenthal and E. Shevach, 1974, J. Exp. Med. 138:1194, by copyright permission of the Rockefeller University Press.] that the TC cell and the virus-infected target cell must share class I molecules encoded by the K or D regions of the MHC. Thus, antigen recognition by CD8 TC cells is class I MHC Spleen cells (containing Tc cells) H–2k target cells H–2k LCM-infected target cells H–2b LCM-infected target cells –51Cr release (no lysis) –51Cr release (no lysis) +51Cr release (lysis) H–2k LCM virus 51Cr FIGURE 8-2 Classic experiment of Zinkernagel and Doherty demonstrating that antigen recognition by TC cells exhibits MHC re￾striction. H-2k mice were primed with the lymphocytic choriomenin￾gitis (LCM) virus to induce cytotoxic T lymphocytes (CTLs) specific for the virus. Spleen cells from this LCM-primed mouse were then added to target cells of different H-2 haplotypes that were intracellu￾larly labeled with 51Cr (black dots) and either infected or not with the LCM virus. CTL-mediated killing of the target cells, as measured by the release of 51Cr into the culture supernatant, occurred only if the target cells were infected with LCM and had the same MHC haplo￾type as the CTLs. [Adapted from P. C. Doherty and R. M. Zinkernagel, 1975, J. Exp. Med. 141:502.] 8536d_ch08_185-199 8/2/02 10:08 AM Page 186 mac79 Mac 79:45_BW:Goldsby et al. / Immunology 5e: