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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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Cytokine levels of MBP-specific TCL stimulated overnight with syngeneic APC. Established lymph node TCL (TCL 2) from mMBP Ac116–immunized B10.A, B10.BR, and (B10.A × B10.BR) F1 mice were restimulated for the fifth time with mMBP Ac1-16 and irradiated splenic APC.  After 24 h, culture supernatants were harvested. Additional cells harvested 3 d after the same fifth restimulation were used for adoptive transfer experiment EAE (see Table 1). Data presented for TNF are means and standard deviations of two bioassays performed on triplicate wells for each sample (n =  6). Data presented for IFN-γ, IL-12, IL-4, and IL-10 are means and standard deviations of triplicate wells for each sample (n = 3). Data presented for  IL-2 are means and standard deviations of two IL-2 bioassays performed in duplicate wells for each sample (n = 4).
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Figure 1: Cytokine levels of MBP-specific TCL stimulated overnight with syngeneic APC. Established lymph node TCL (TCL 2) from mMBP Ac116–immunized B10.A, B10.BR, and (B10.A × B10.BR) F1 mice were restimulated for the fifth time with mMBP Ac1-16 and irradiated splenic APC. After 24 h, culture supernatants were harvested. Additional cells harvested 3 d after the same fifth restimulation were used for adoptive transfer experiment EAE (see Table 1). Data presented for TNF are means and standard deviations of two bioassays performed on triplicate wells for each sample (n = 6). Data presented for IFN-γ, IL-12, IL-4, and IL-10 are means and standard deviations of triplicate wells for each sample (n = 3). Data presented for IL-2 are means and standard deviations of two IL-2 bioassays performed in duplicate wells for each sample (n = 4).

Mentions: In contrast, comparison of the cytokine profiles of B10.A and B10.BR TCL revealed differences which correlated with their encephalitogenicity. Data for the TCL 2 lines are shown in Fig. 1. The B10.A TCL stimulated with peptide and B10.A splenic APC displayed the Th2 phenotype, expressing very low levels of IFN-γ, IL-2, and TNF-α and high levels of IL-4 and IL-10. Levels of Th1-promoting IL-12 (made by APC) were undetectable. In contrast, the B10.BR TCL displayed the Th1 phenotype, secreting very little IL-4 and IL-10, and high levels of IFN-γ, IL-2, and TNF-α. IL-12 levels were also significantly higher. TNF-α ELISA results (not shown) were nearly identical (within 10%) to the TNF bioassay results, and as TNF biological activity was totally blocked with the anti–TNF-α mAb TN3-19.12 (see below), all of the TNF activity detected in these TCL supernatants appeared to be TNF-α. Supernatants from the (B10.A × B10.BR) F1 TCL stimulated with peptide and F1 APC contained intermediate amounts of the tested cytokines, but the levels were closer to those from B10.A than B10.BR TCL. Thus, TCL from B10.A mice produced a Th2, low TNF-α response, T cells from B10.BR produced a Th1, high TNF-α response, and the Th2, low TNF-α phenotype was dominant in the (B10.A × B10.BR) F1 T cell cultures. These results indicated that genetic difference between B10.A and B10.BR controls a regulator of Th cytokines.


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)

Cytokine levels of MBP-specific TCL stimulated overnight with syngeneic APC. Established lymph node TCL (TCL 2) from mMBP Ac116–immunized B10.A, B10.BR, and (B10.A × B10.BR) F1 mice were restimulated for the fifth time with mMBP Ac1-16 and irradiated splenic APC.  After 24 h, culture supernatants were harvested. Additional cells harvested 3 d after the same fifth restimulation were used for adoptive transfer experiment EAE (see Table 1). Data presented for TNF are means and standard deviations of two bioassays performed on triplicate wells for each sample (n =  6). Data presented for IFN-γ, IL-12, IL-4, and IL-10 are means and standard deviations of triplicate wells for each sample (n = 3). Data presented for  IL-2 are means and standard deviations of two IL-2 bioassays performed in duplicate wells for each sample (n = 4).
© Copyright Policy
Related In: Results  -  Collection

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Figure 1: Cytokine levels of MBP-specific TCL stimulated overnight with syngeneic APC. Established lymph node TCL (TCL 2) from mMBP Ac116–immunized B10.A, B10.BR, and (B10.A × B10.BR) F1 mice were restimulated for the fifth time with mMBP Ac1-16 and irradiated splenic APC. After 24 h, culture supernatants were harvested. Additional cells harvested 3 d after the same fifth restimulation were used for adoptive transfer experiment EAE (see Table 1). Data presented for TNF are means and standard deviations of two bioassays performed on triplicate wells for each sample (n = 6). Data presented for IFN-γ, IL-12, IL-4, and IL-10 are means and standard deviations of triplicate wells for each sample (n = 3). Data presented for IL-2 are means and standard deviations of two IL-2 bioassays performed in duplicate wells for each sample (n = 4).
Mentions: In contrast, comparison of the cytokine profiles of B10.A and B10.BR TCL revealed differences which correlated with their encephalitogenicity. Data for the TCL 2 lines are shown in Fig. 1. The B10.A TCL stimulated with peptide and B10.A splenic APC displayed the Th2 phenotype, expressing very low levels of IFN-γ, IL-2, and TNF-α and high levels of IL-4 and IL-10. Levels of Th1-promoting IL-12 (made by APC) were undetectable. In contrast, the B10.BR TCL displayed the Th1 phenotype, secreting very little IL-4 and IL-10, and high levels of IFN-γ, IL-2, and TNF-α. IL-12 levels were also significantly higher. TNF-α ELISA results (not shown) were nearly identical (within 10%) to the TNF bioassay results, and as TNF biological activity was totally blocked with the anti–TNF-α mAb TN3-19.12 (see below), all of the TNF activity detected in these TCL supernatants appeared to be TNF-α. Supernatants from the (B10.A × B10.BR) F1 TCL stimulated with peptide and F1 APC contained intermediate amounts of the tested cytokines, but the levels were closer to those from B10.A than B10.BR TCL. Thus, TCL from B10.A mice produced a Th2, low TNF-α response, T cells from B10.BR produced a Th1, high TNF-α response, and the Th2, low TNF-α phenotype was dominant in the (B10.A × B10.BR) F1 T cell cultures. These results indicated that genetic difference between B10.A and B10.BR controls a regulator of Th cytokines.

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