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Immunity | 1996

IL-12-Deficient Mice Are Defective in IFNγ Production and Type 1 Cytokine Responses

Jeanne Magram; Suzanne E. Connaughton; Rajeev R. Warrier; Daisy Carvajal; Chang-You Wu; Jessica Ferrante; Colin L. Stewart; Ulla Sarmiento; Denise A. Faherty; Maurice K. Gately

Abstract IL-12 is a cytokine that can exert regulatory effects on T and NK cells and promote Th1 responses. To delineate further the physiologic role of IL-12 in immunity, mice deficient for this cytokine were generated. IL-12-deficient mice were impaired but not completely lacking in the ability to produce IFNγ following endotoxin administration and to mount a Th1 response in vivo, as measured by antigen-induced IFNγ secretion by immune lymph node cells in vitro. In contrast, secretion of IL-4 was enhanced, while proliferation and secretion of IL-2 and IL-10 were normal following antigen stimulation. DTH responses were significantly reduced in IL-12-deficient mice, but no defect in allogeneic CTL responses was observed. These results indicate that IL-12 plays an essential role in regulating IFNγ production and in facilitating normal DTH responses. However, other phenomena associated with Th1 responses and cell-mediated immunity, i.e., IL-2 secretion and CTL generation, were not compromised in the absence of IL-12.


Annals of the New York Academy of Sciences | 1996

Interleukin-12 Antagonist Activity of Mouse Interleukin-12 p40 Homodimer in Vitro and in Vivo

Maurice K. Gately; Daisy Carvajal; Suzanne E. Connaughton; Silke Gillessen; Rajeev R. Warrier; Kenneth Kolinsky; Victoria L. Wilkinson; Cynthia M. Dwyer; George F. Higgins; Frank John Podlaski; Denise A. Faherty; Philip C. Familletti; Alvin S. Stern; David H. Presky

Mo(p40)2 is a potent IL-12 antagonist that interacts strongly with the beta 1 subunit of the IL-12R to block binding of moIL-12 to the high-affinity mouse IL-12R. Mo(p40)2, alone or in synergy with the 2B5 mAb specific for the moIL-12 heterodimer, blocked IL-12-induced responses in vitro, Mo(p40)2 was thus used alone or with 2B5 mAb to examine the role of IL-12 in vivo, Mo(p40)2 caused a dose-dependent inhibition of both the rise in serum IFN-gamma levels in mice injected with endotoxin and the Th1-like response to immunization with KLH. Treatment with mo(p40)2 plus 2B5 anti-moIL-12 mAb also suppressed DTH responses to methylated bovine serum albumin but not specific allogeneic CTL responses in vivo. In each of these models, responses seen in mice treated with mo(p40)2 +/- 2B5 anti-moIL-12 mAb were similar to those observed in IL-12 knockout mice. Thus, mo(p40)2 can act as a potent IL-12 antagonist in vivo, as well as in vitro, and is currently being used to investigate the role of IL-12 in the pathogenesis of some Th1-associated autoimmune disorders in mice.


Annals of the New York Academy of Sciences | 1996

IL-12-Deficient Mice Are Defective but Not Devoid of Type 1 Cytokine Responses

Jeanne Magram; Jessica Sfarra; Suzanne E. Connaughton; Denise A. Faherty; Rajeev R. Warrier; Daisy Carvajal; Chang-You Wu; Colin L. Stewart; Ulla Sarmiento; Maurice K. Gately

Interleukin-12 (IL-12) has been described as a pivotal molecule in the immune response based in part on its ability to influence the differentiation of T helper (Th) cells into a type 1 (Th1) phenotype. This event is crucial in that appropriate differentiation of naive T cells can determine susceptibility or resistance to given pathogens by influencing the balance between cellular and humoral immunity. In order to further delineate the role of IL-12 in the immune response, we generated mice deficient for this cytokine. IL-12 knockout mice were viable, fully fertile, and displayed no obvious developmental abnormalities. Upon immunological analysis, these mice demonstrated an impaired ability to effect a Th1 response as well as an impaired ability to produce interferon-gamma in response to endotoxin in vivo. These data establish an essential role for IL-12 in the generation of optimal Th1 responses in vivo, but weak responses can occur independently of IL-12.


Journal of Immunological Methods | 1996

Characterization of anti-mouse IL-12 monoclonal antibodies and measurement of mouse IL-12 by ELISA.

Victoria L. Wilkinson; Rajeev R. Warrier; Theresa Truitt; Perla Nunes; Maurice K. Gately; David H. Presky

Enzyme-linked immunosorbent assays (ELISAs) capable of quantitatively measuring pg/ml amounts of mouse IL-12 (moIL-12) were developed as an alternative to the current bioassay procedure used for the measurement of moIL-12. A panel of 40 rat anti-moIL-12 monoclonal antibodies were identified and tested for their ability to bind 125I-moIL-12. Two of the MAbs, 2B5 and 9A5, were able to capture 125I-moIL-12 in the presence of unlabelled moIL-12 p35 and moIL-12 p40, suggesting specificity for the moIL-12 p75 heterodimer. Western blot analysis confirmed that MAb 9A5 specifically recognized only moIL-12 p75. Using MAb 9A5, and an additional anti-moIL-12 p40 MAb 5D9, we developed quantitative ELISAs for the specific detection of moIL-12 p75 and p40, respectively. These ELISAs detect moIL-12 with a sensitivity of 40 pg/ml. Whereas the p40 ELISA detected three forms of moIL-12 (p40 monomer, p40 homodimer, and the heterodimer), the p75 ELISA only detected moIL-12 heterodimer. Neither of these assays crossreacted with a panel of additional cytokines. The levels of moIL-12 measured by the p75 ELISA and the bioassay were directly compared and found to correlate well. Therefore, the p75 ELISA represents an alternative to the bioassay for the measurement of moIL-12.


Journal of Experimental Medicine | 1993

Antitumor and Antimetastatic Activity of Interleukin 12 against Murine Tumors

Michael J. Brunda; Leopoldo Luistro; Rajeev R. Warrier; Rosemary B. Wright; Brian R. Hubbard; Molly Murphy; Stanley F. Wolf; Maurice K. Gately


International Immunology | 1994

Administration of recombinant IL-12 to normal mice enhances cytolytic lymphocyte activity and induces production of IFN-γ in vivo

Maurice K. Gately; Rajeev R. Warrier; Sharmila Honasoge; Daisy Carvajal; Denise A. Faherty; Suzanne E. Connaughton; Timothy D. Anderson; Ulla Sarmiento; Brian R. Hubbard; Molly Murphy


European Journal of Immunology | 1996

Reduced incidence and severity of collagen‐induced arthritis in interleukin‐12‐deficient mice

Kim W. McIntyre; David J. Shuster; Kathleen M. Gillooly; Rajeev R. Warrier; Suzanne E. Connaughton; Leroy B. Hall; Lawrence H. Arp; Maurice K. Gately; Jeanne Magram


European Journal of Immunology | 1997

Regulation of interleukin‐12 receptor β1 chain expression and interleukin‐12 binding by human peripheral blood mononuclear cells

Chang-You Wu; Rajeev R. Warrier; Xin Wang; David H. Presky; Maurice K. Gately


European Journal of Immunology | 1996

Biological function and distribution of human interleukin-12 receptor β chain

Chang-You Wu; Rajeev R. Warrier; Daisy Carvajal; Anne O. Chua; Lisa J. Minetti; Richard Anthony Chizzonite; Patricia K. A. Mongini; Alvin S. Stern; Ueli Gubler; David H. Presky; Maurice K. Gately


Journal of Interferon and Cytokine Research | 1995

Administration of Recombinant Interleukin-12 to Mice Suppresses Hematopoiesis in the Bone Marrow but Enhances Hematopoiesis in the Spleen

Nadine Tare; Sharon Bowen; Rajeev R. Warrier; Daisy Carvajal; William R. Benjamin; Julia H. Riley; Timothy D. Anderson; Maurice K. Gately

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Colin L. Stewart

Roche Institute of Molecular Biology

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