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Novel genetic regulation of T helper 1 (Th1)/Th2 cytokine production and encephalitogenicity in inbred mouse strains.

Conboy IM, DeKruyff RH, Tate KM, Cao ZA, Moore TA, Umetsu DT, Jones PP - J. Exp. Med. (1997)

Bottom Line: A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone.The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL.The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Stanford University, California 94305-5020, USA.

ABSTRACT
Development of T helper cell (Th)1 or Th2 cytokine responses is essential for effector and regulatory functions of T helper cells. We have compared cytokine profiles of myelin basic protein (MBP) Ac1-16 peptide-specific T helper cells from inbred mouse strains expressing identical k haplotype-derived MHC class II molecules B10.A and B10.BR, B10.BR T cell lines (TCL) produced Th1 cytokines (including high levels of TNF-alpha) and induced experimental autoimmune encephalomyelitis after adoptive transfer. In contrast, B10.A TCL produced Th2 cytokines (including low levels of TNF-alpha) and were poorly encephalitogenic. The contributions of the genetic origin of the T cells and the APC were explored. Serial restimulations of the B10.BR TCL with B10.A or (B10.A x B10.BR) F1 splenic antigen presenting cells (APC) during the establishment of TCL markedly reduced both Th1 cytokine production and encephalitogenicity. In addition, a single restimulation with B10. A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone. These studies suggest that B10.A and B10.BR APC differ in their ability to stimulate IFN-gamma and TNF-alpha production by mature Th1 cells and also influence their Th1/Th2 commitment in vivo. The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL. The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2. Thus, genetic difference(s) between B10.A and B10.BR APC appear(s) to control the production or activity of a novel soluble cytokine regulatory factor that influences Th1/Th2 commitment and controls production of IFN-gamma and TNF-alpha by mature Th1 cells.

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Cytokines produced by TCL (TCL 3) from B10.A and  B10.BR mice are influenced by the source of APC (B10.A, B10.BR, or  F1) used for in vitro restimulations. (A–D) After the third, fourth, fifth,  ninth, and tenth restimulations of TCL with mMBP Ac1-16 and irradiated splenic APC of the indicated origin, T cells were restimulated for 24 h  with peptide and APC of the same origin. Adoptive transfer of EAE was  performed after the ninth restimulation (see Table 2). Supernatant fluids  were analyzed for cytokines. In Fig. 2 D, the points for the B10.BR  TCL-3–B10.A APC are 37 ng/ml of IL-10 after the ninth restimulation  and 45 ng/ml of IL-10 after the tenth restimulation. Data presented are  average of duplicate wells for each sample. (E) At the ninth restimulation,  TCL 3 described for A–D were restimulated for 24 h with peptide and  APC of the same (indicated) origin. Supernatant fluids depleted of IL-4  were analyzed for IL-2 levels, using the growth factor dependent cell line  HT-2. Data presented are means and standard deviations of four wells for  each sample (n = 4).
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Figure 2: Cytokines produced by TCL (TCL 3) from B10.A and B10.BR mice are influenced by the source of APC (B10.A, B10.BR, or F1) used for in vitro restimulations. (A–D) After the third, fourth, fifth, ninth, and tenth restimulations of TCL with mMBP Ac1-16 and irradiated splenic APC of the indicated origin, T cells were restimulated for 24 h with peptide and APC of the same origin. Adoptive transfer of EAE was performed after the ninth restimulation (see Table 2). Supernatant fluids were analyzed for cytokines. In Fig. 2 D, the points for the B10.BR TCL-3–B10.A APC are 37 ng/ml of IL-10 after the ninth restimulation and 45 ng/ml of IL-10 after the tenth restimulation. Data presented are average of duplicate wells for each sample. (E) At the ninth restimulation, TCL 3 described for A–D were restimulated for 24 h with peptide and APC of the same (indicated) origin. Supernatant fluids depleted of IL-4 were analyzed for IL-2 levels, using the growth factor dependent cell line HT-2. Data presented are means and standard deviations of four wells for each sample (n = 4).

Mentions: The cytokine profiles of the third set (TCL 3) of MBP-specific T cell lines serially restimulated with splenic APC of varying genetic origins were also examined. The cytokine responses of the six TCL were determined after the third, fourth, fifth, ninth, and tenth restimulations. We found that the genetic origin of the splenic APC used for restimulations greatly influenced the cytokine profile of the B10.BR TCL. Thus, the Th1, high TNF-α phenotype of the B10.BR TCL was maintained only by syngeneic B10.BR splenic APC; very little IFN-γ, TNF-α, or IL-2 was produced by the B10.BR TCL serially restimulated with F1 or B10.A APC (Fig. 2). For IL-4, all three of the B10.A TCL had higher levels than the B10.BR TCL (Fig. 2 C), and B10.BR T cells maintained on B10.BR APC produced the lowest IL-4 levels. IL-10, B10.A, and F1 APC, but not B10.BR APC, supported levels similar to those produced by B10.A TCL (Fig. 2 D). A Th2, low TNF-α profile was seen for all B10.A TCL irrespective of the source of APC.


Novel genetic regulation of T helper 1 (Th1)/Th2 cytokine production and encephalitogenicity in inbred mouse strains.

Conboy IM, DeKruyff RH, Tate KM, Cao ZA, Moore TA, Umetsu DT, Jones PP - J. Exp. Med. (1997)

Cytokines produced by TCL (TCL 3) from B10.A and  B10.BR mice are influenced by the source of APC (B10.A, B10.BR, or  F1) used for in vitro restimulations. (A–D) After the third, fourth, fifth,  ninth, and tenth restimulations of TCL with mMBP Ac1-16 and irradiated splenic APC of the indicated origin, T cells were restimulated for 24 h  with peptide and APC of the same origin. Adoptive transfer of EAE was  performed after the ninth restimulation (see Table 2). Supernatant fluids  were analyzed for cytokines. In Fig. 2 D, the points for the B10.BR  TCL-3–B10.A APC are 37 ng/ml of IL-10 after the ninth restimulation  and 45 ng/ml of IL-10 after the tenth restimulation. Data presented are  average of duplicate wells for each sample. (E) At the ninth restimulation,  TCL 3 described for A–D were restimulated for 24 h with peptide and  APC of the same (indicated) origin. Supernatant fluids depleted of IL-4  were analyzed for IL-2 levels, using the growth factor dependent cell line  HT-2. Data presented are means and standard deviations of four wells for  each sample (n = 4).
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Figure 2: Cytokines produced by TCL (TCL 3) from B10.A and B10.BR mice are influenced by the source of APC (B10.A, B10.BR, or F1) used for in vitro restimulations. (A–D) After the third, fourth, fifth, ninth, and tenth restimulations of TCL with mMBP Ac1-16 and irradiated splenic APC of the indicated origin, T cells were restimulated for 24 h with peptide and APC of the same origin. Adoptive transfer of EAE was performed after the ninth restimulation (see Table 2). Supernatant fluids were analyzed for cytokines. In Fig. 2 D, the points for the B10.BR TCL-3–B10.A APC are 37 ng/ml of IL-10 after the ninth restimulation and 45 ng/ml of IL-10 after the tenth restimulation. Data presented are average of duplicate wells for each sample. (E) At the ninth restimulation, TCL 3 described for A–D were restimulated for 24 h with peptide and APC of the same (indicated) origin. Supernatant fluids depleted of IL-4 were analyzed for IL-2 levels, using the growth factor dependent cell line HT-2. Data presented are means and standard deviations of four wells for each sample (n = 4).
Mentions: The cytokine profiles of the third set (TCL 3) of MBP-specific T cell lines serially restimulated with splenic APC of varying genetic origins were also examined. The cytokine responses of the six TCL were determined after the third, fourth, fifth, ninth, and tenth restimulations. We found that the genetic origin of the splenic APC used for restimulations greatly influenced the cytokine profile of the B10.BR TCL. Thus, the Th1, high TNF-α phenotype of the B10.BR TCL was maintained only by syngeneic B10.BR splenic APC; very little IFN-γ, TNF-α, or IL-2 was produced by the B10.BR TCL serially restimulated with F1 or B10.A APC (Fig. 2). For IL-4, all three of the B10.A TCL had higher levels than the B10.BR TCL (Fig. 2 C), and B10.BR T cells maintained on B10.BR APC produced the lowest IL-4 levels. IL-10, B10.A, and F1 APC, but not B10.BR APC, supported levels similar to those produced by B10.A TCL (Fig. 2 D). A Th2, low TNF-α profile was seen for all B10.A TCL irrespective of the source of APC.

Bottom Line: A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone.The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL.The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Stanford University, California 94305-5020, USA.

ABSTRACT
Development of T helper cell (Th)1 or Th2 cytokine responses is essential for effector and regulatory functions of T helper cells. We have compared cytokine profiles of myelin basic protein (MBP) Ac1-16 peptide-specific T helper cells from inbred mouse strains expressing identical k haplotype-derived MHC class II molecules B10.A and B10.BR, B10.BR T cell lines (TCL) produced Th1 cytokines (including high levels of TNF-alpha) and induced experimental autoimmune encephalomyelitis after adoptive transfer. In contrast, B10.A TCL produced Th2 cytokines (including low levels of TNF-alpha) and were poorly encephalitogenic. The contributions of the genetic origin of the T cells and the APC were explored. Serial restimulations of the B10.BR TCL with B10.A or (B10.A x B10.BR) F1 splenic antigen presenting cells (APC) during the establishment of TCL markedly reduced both Th1 cytokine production and encephalitogenicity. In addition, a single restimulation with B10. A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone. These studies suggest that B10.A and B10.BR APC differ in their ability to stimulate IFN-gamma and TNF-alpha production by mature Th1 cells and also influence their Th1/Th2 commitment in vivo. The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL. The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2. Thus, genetic difference(s) between B10.A and B10.BR APC appear(s) to control the production or activity of a novel soluble cytokine regulatory factor that influences Th1/Th2 commitment and controls production of IFN-gamma and TNF-alpha by mature Th1 cells.

Show MeSH
Related in: MedlinePlus