1. Volume 121, Issue 4, Pages 931-939 (October 2001)
Immunotherapy directed against α-fetoprotein results in autoimmune liver disease during liver regeneration in mice 
Michael Geissler, Leonhard Mohr, Robert Weth, Gabriele Köhler, Christian F. Grimm, Tim U. Krohne, Fritz Von Weizsäcker, Hubert E. Blum 
Gastroenterology 
Volume 121, Issue 4, Pages (October 2001)
DOI: /gast
Copyright © 2001 American Gastroenterological Association Terms and Conditions

2. Fig. 1
CTL activity in AFP-DNA immunized C57BL/6 mice. (A) CD8+ T cells derived from AFP- or mock-DNA immunized mice (n = 4 in each group) were stimulated for 26 hours with rVV-mAFP or rVV-pSC11, or left unstimulated. Subsequently, IFN-γ ELISPOT assays were performed. The spots in each well were counted under a microscope, and the values are expressed as numbers of spot-forming cells relative to the number of spleen cells added to each well at the start of the culture. (B and C) Single spleen cell suspensions derived from (B) pcD/3-mAFP– or (C) pSec-mAFP–immunized mice (n = 4 in each group) were assayed after in vitro stimulation with rVV-mAFP–infected syngeneic spleen cells for 8 days. The effector cells were then tested against MC57G target cells infected with the indicated vaccinia viruses in a 51Cr-release assay at different E:T ratios. Values represent means of triplicate determinations.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

3. Fig. 2
CTL activity in AFP-DNA immunized MHC II knockout mice. (A) CD8+ T cells derived from AFP- or mock-DNA immunized MHC II knockout mice (n = 3 in each group) were stimulated for 20 hours with rVV-mAFP or rVV-pSC11, or left unstimulated. Subsequently, IFN-γ ELISPOT assays were performed. (B and C) Single spleen cell suspensions derived from (B) pcD/3-mAFP or (C) pSec-mAFP immunized MHC II knockout mice (n = 3) were assayed after in vitro stimulation with rVV-mAFP–infected syngeneic spleen cells for 7 days. The effector cells were then tested against MC57G target cells infected with the different vaccinia viruses in a 51Cr-release assay at the E:T ratio indicated.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Time course of AFP-specific CTL-p frequency and transaminases (ALT). (A) Time course of AFP-specific CD8+ T-cell precursor frequency after immunization of C57BL/6 mice with AFP-DNA expression plasmids at day 1 and 14. Spleen cells derived from AFP- or mock-DNA immunized mice (n = 4 in each group) were stimulated for 24 hours with rVV-mAFP. Subsequently, IFN-γ ELISPOT assays were performed. (B) Time course of ALT levels and histologic examination of the liver of AFP-DNA–immunized mice for the presence of lymphocytic infiltrates (−, no infiltrates; (+), few scattered microfocal infiltrates; and +, microfocal and focal infiltrates).
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Histochemical analysis of livers. Bars in pictures A–F correspond to 0.2 mm, and in inserts of A, B, C, D, and F to 0.03 mm. (A) Mock-DNA, C57BL/6 mice, 30 hours after partial hepatectomy. (Insert) BrdU-staining, bar corresponds to mm. (B) AFP-DNA, C57B46 mice 6 days after the first immunization. (C) AFP-DNA, C57BL/6 mice, 46 hours after partial hepatectomy. (D) AFP-DNA, C57BL/6 mice, 120 hours after partial hepatectomy. (F) AFP-DNA immunized CD4+ T cell–depleted mice, 5 days after the second immunization (day 19).
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Experimental design of the study. C57L/6 and C3H mice were DNA-immunized at days 1 and 14. Five days later, partial hepatectomy was performed and mice were monitored by serially measuring serum ALT, by serial histologic examinations of the liver, and by the determination of the time course of CTL responses directed against AFP. Identification of pathogenic immune effectors was done by using MHC class I knockout mice and in vivo T-cell depletion experiments.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Time course of AFP-specific CTL-p frequency and transaminases (ALT) in CD4+ and CD8+ T cell–depleted mice. (A) C57BL/6 mice were immunized with AFP- or mock-DNA expression plasmids at days 1 and 14. Spleen cells derived from AFP- or mock-DNA immunized mice (n = 2 in each group) were stimulated for 20 hours with rVV-mAFP. Subsequently, IFN-γ ELISPOT assays were performed at the indicated time points. CD4+ and CD8+ T-cell subpopulations were depleted by IP injection of purified hybridoma supernatant. A total of 1 mg per mouse per injection of anti-CD8 or anti-CD4 was injected on days 14, 16, 18, 21, and 23 after the first immunization. (B) Time course of ALT levels in AFP-DNA immunized T cell–depleted mice (n = 2 in each group).
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

8. Fig. 7
ALT levels and AFP-specific CTL-p frequency after partial hepatectomy. Mock- (n = 13) or AFP-DNA (n = 17) immunized (days 1 and 14) C57BL/6 were partially hepatectomized at day 19 after the first immunization (corresponds to 0 hours in this figure; n = 7 and 10, respectively) or left without hepatectomy (n = 6 and 7, respectively). Similarly, C3H mice were immunized with mock- (n = 9) or AFP-DNA (n = 10), and partial hepatectomy was performed in 7 mock- and 8 AFP-DNA immunized mice, respectively. (A) ALT levels in C57BL/6 and C3H mice and (B) CTL-p frequencies in C57BL/6 mice were serially measured.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

9. Fig. 8
Identification of autoimmune mediators. Time course of ALT levels in naive or AFP-DNA (days 1 and 14) immunized MHC I knockout mice after partial hepatectomy performed at day 19 (corresponds to 0 hours).
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions