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Novel antibodies that selectively block mouse IL‐12 enable the re‐evaluation of the role of IL‐12 in immune protection and pathology

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Melanie Gaignage at Université Catholique de Louvain - UCLouvain
Melanie Gaignage
Catherine Uyttenhove at Université Catholique de Louvain - UCLouvain
Catherine Uyttenhove
Lindsay L. Jones
Lindsay L. Jones
Lindsay L. Jones
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Christophe Bourdeaux
Christophe Bourdeaux
Christophe Bourdeaux
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Abstract and Figures

The dimeric cytokine IL‐12 is important in the control of various infections but also contributes to the pathology of certain diseases making it a potential target for therapy. However, its specific inhibition with antibodies is complicated by the fact that its two subunits are present in other cytokines: p40 in IL‐23 and p35 in IL‐35. This has led to erroneous conclusions like the alleged implication of IL‐12 in experimental autoimmune encephalomyelitis (EAE). Here, we report the development of a mouse anti‐mouse IL‐12 vaccine and the production of monoclonal antibodies (mAbs) that do not react with p40 or p35 (in IL‐35) but specifically recognize and functionally inhibit the IL‐12 heterodimer. Using one of these mAbs, MM12A1.6, that strongly inhibited IFNγ production and LPS‐induced septic shock after viral infection, we demonstrate the critical role played by IL‐12 in the rejection of male skin graft by female C57Bl/6 syngeneic recipients and in the clearance of an immunogenic mastocytoma tumor variant by DBA/2 mice, but not in a parent to F1 immune aggression model nor in MOG‐induced EAE, which was clearly prevented by anti‐p40 mAb C17.8. Given this selective inhibition of IL‐12, these mAbs provide new options for reassessing IL‐12 function in vivo. This article is protected by copyright. All rights reserved
MM12 Abs specifically recognize IL‐12p35‐p40 but not IL‐35, p40, or IL‐23. (A) B6 mice immunized with mouse IL‐12 conjugated to OVAglu polymers were used to produce anti‐IL‐12 hybridomas that were designated MM12. Binding of IL‐12 to six such mAbs (A1.6, A2.23, A2.36, A2.69, A2.103, and A2.107), anti‐IL‐12p35 Nter peptide Ab MMp35.8G7 (8G7), and anti‐p40 Ab C17.8 is shown here. Briefly, ELISA plates coated with avidin (5 μg/mL) or BSA were incubated with biotinylated IL‐12 at 200 ng/mL for 1 h, washed and then incubated with various concentrations of Abs for 2 h. Bound Ab was detected with HRP‐conjugated goat anti‐mouse Ig followed by TMB and absorbance reading at 450 nm. nM concentrations for half‐ maximal binding are indicated (nM 50%). Data are representative of two independent experiments with technical duplicates per experiment. (B) Binding of biotinylated IL‐12, p40, IL‐35 (p28‐human Ig‐Ebi3), and IL‐23, all at 200 ng/mL, to immobilized Abs or BSA detected with avidin‐HRP (mean of triplicates ± SEM). Data are representative of two independent experiments with technical triplicates per experiment, where IL‐35 was detected with goat anti‐human IgG‐HRP. (C) Single‐chain constructs of IL‐12 (p40‐linker‐p35) and IL‐35 (Ebi3‐linker‐p35) were cloned into pCT302 yeast display vector before being transformed into S. cerevisiae yeast display strain EBY100. Yeast cells expressing IL‐12 or IL‐35 were incubated with the indicated Ab followed by anti‐mouse Alexa 647 and flow cytometry analysis. Uninduced IL‐12 yeast grown in the absence of galactose and lacking surface expression were stained as a negative control. Data representative of two experiments performed in monoplicate.
MM12 Abs specifically recognize IL‐12p35‐p40 but not IL‐35, p40, or IL‐23. (A) B6 mice immunized with mouse IL‐12 conjugated to OVAglu polymers were used to produce anti‐IL‐12 hybridomas that were designated MM12. Binding of IL‐12 to six such mAbs (A1.6, A2.23, A2.36, A2.69, A2.103, and A2.107), anti‐IL‐12p35 Nter peptide Ab MMp35.8G7 (8G7), and anti‐p40 Ab C17.8 is shown here. Briefly, ELISA plates coated with avidin (5 μg/mL) or BSA were incubated with biotinylated IL‐12 at 200 ng/mL for 1 h, washed and then incubated with various concentrations of Abs for 2 h. Bound Ab was detected with HRP‐conjugated goat anti‐mouse Ig followed by TMB and absorbance reading at 450 nm. nM concentrations for half‐ maximal binding are indicated (nM 50%). Data are representative of two independent experiments with technical duplicates per experiment. (B) Binding of biotinylated IL‐12, p40, IL‐35 (p28‐human Ig‐Ebi3), and IL‐23, all at 200 ng/mL, to immobilized Abs or BSA detected with avidin‐HRP (mean of triplicates ± SEM). Data are representative of two independent experiments with technical triplicates per experiment, where IL‐35 was detected with goat anti‐human IgG‐HRP. (C) Single‐chain constructs of IL‐12 (p40‐linker‐p35) and IL‐35 (Ebi3‐linker‐p35) were cloned into pCT302 yeast display vector before being transformed into S. cerevisiae yeast display strain EBY100. Yeast cells expressing IL‐12 or IL‐35 were incubated with the indicated Ab followed by anti‐mouse Alexa 647 and flow cytometry analysis. Uninduced IL‐12 yeast grown in the absence of galactose and lacking surface expression were stained as a negative control. Data representative of two experiments performed in monoplicate.
… 
MM12 mAbs recognize a limited number of epitopes on IL‐12p35‐p40 heterodimer that correlate with their inhibitory activity. (A) B6 mice immunized with mouse IL‐12 conjugated to OVAglu polymers were used to produce anti‐IL‐12 hybridomas that were designated MM12. Inhibition of IL‐12‐induced proliferation of Ba/F3 mIL‐12R cells by MM12 Abs, C17.8, and a control IgG2a by serial dilutions of the Abs and representative of two distinct experiments. Mean ± SD are computed from technical duplicate in each experiment. IC50 calculated for each Ab are indicated. (B) Competition of MM12 Ab for binding to IL‐12: biotinylated IL‐12 (200 ng/mL) was incubated with 20 μg/mL of each Ab overnight before transferring the mix to ELISA plates coated with the different MM12 Abs. The positive control (Control) is a well with coated Ab + biotinylated IL‐12 only and negative control (Blank) is a well without coated Ab and incubated with the mix of biotinylated IL‐12 + mAbs. After 1 h incubation at 37°C and washing, plate‐bound biotinylated IL‐12 was measured by 1 h incubation with avidin‐HRP followed by TMB. The results correspond to duplicate samples and are representative of two independent experiments.
MM12 mAbs recognize a limited number of epitopes on IL‐12p35‐p40 heterodimer that correlate with their inhibitory activity. (A) B6 mice immunized with mouse IL‐12 conjugated to OVAglu polymers were used to produce anti‐IL‐12 hybridomas that were designated MM12. Inhibition of IL‐12‐induced proliferation of Ba/F3 mIL‐12R cells by MM12 Abs, C17.8, and a control IgG2a by serial dilutions of the Abs and representative of two distinct experiments. Mean ± SD are computed from technical duplicate in each experiment. IC50 calculated for each Ab are indicated. (B) Competition of MM12 Ab for binding to IL‐12: biotinylated IL‐12 (200 ng/mL) was incubated with 20 μg/mL of each Ab overnight before transferring the mix to ELISA plates coated with the different MM12 Abs. The positive control (Control) is a well with coated Ab + biotinylated IL‐12 only and negative control (Blank) is a well without coated Ab and incubated with the mix of biotinylated IL‐12 + mAbs. After 1 h incubation at 37°C and washing, plate‐bound biotinylated IL‐12 was measured by 1 h incubation with avidin‐HRP followed by TMB. The results correspond to duplicate samples and are representative of two independent experiments.
… 
MM12A1.6 prevents viral‐induced IFN‐γ production and impairs LPS‐induced septic shock in IFNAR‐1−/− and WT 129/Sv mice. (A) Schematic representation illustrates the experimental protocol. IFNAR‐1−/− and WT 129/Sv mice were injected or not with 0.5 mg MM12A1.6 mAb or control IgG2a AH9R3A mAb i.p. 4 h before infection with LDV. (B) Plasma IFN‐γ concentrations were measured by ELISA 24 h after infection (five mice per group). Data are representative of two experiments. Each data point represents the value measured in individual mice with n = 5 for LDV + isotype – WT; n = 6 for LDV + MM12A1.6 – WT; n = 5 for LDV + isotype – IFNAR‐1−/−; n = 5 for LDV + MM12A1.6 − IFNAR‐1−/−, n = 4 for untreated WT and n = 5 for untreated IFNAR‐1−/−. Difference between MM12A1.6 and control LDV was calculated by Mann–Whitney unpaired t‐test (**p < 0.01) and error bars represent mean ± SEM. (C) The same experiment was performed as in B but 10 μg LPS was administered 24 h after LDV and survival was monitored. Overall survivals of two experiments are depicted with a total of ten mice for LDV + isotype – WT; ten mice for LDV + MM12A1.6 – WT; nine mice for LDV + isotype – IFNAR‐1−/−; ten mice for LDV + MM12A1.6 − IFNAR‐1−/− (**p < 0.01 and ***p < 0.001 by the log‐rank test).
MM12A1.6 prevents viral‐induced IFN‐γ production and impairs LPS‐induced septic shock in IFNAR‐1−/− and WT 129/Sv mice. (A) Schematic representation illustrates the experimental protocol. IFNAR‐1−/− and WT 129/Sv mice were injected or not with 0.5 mg MM12A1.6 mAb or control IgG2a AH9R3A mAb i.p. 4 h before infection with LDV. (B) Plasma IFN‐γ concentrations were measured by ELISA 24 h after infection (five mice per group). Data are representative of two experiments. Each data point represents the value measured in individual mice with n = 5 for LDV + isotype – WT; n = 6 for LDV + MM12A1.6 – WT; n = 5 for LDV + isotype – IFNAR‐1−/−; n = 5 for LDV + MM12A1.6 − IFNAR‐1−/−, n = 4 for untreated WT and n = 5 for untreated IFNAR‐1−/−. Difference between MM12A1.6 and control LDV was calculated by Mann–Whitney unpaired t‐test (**p < 0.01) and error bars represent mean ± SEM. (C) The same experiment was performed as in B but 10 μg LPS was administered 24 h after LDV and survival was monitored. Overall survivals of two experiments are depicted with a total of ten mice for LDV + isotype – WT; ten mice for LDV + MM12A1.6 – WT; nine mice for LDV + isotype – IFNAR‐1−/−; ten mice for LDV + MM12A1.6 − IFNAR‐1−/− (**p < 0.01 and ***p < 0.001 by the log‐rank test).
… 
MM12A1.6 impairs rejection of an immunogenic mouse P815 tumor variant. (A) Schematic representation illustrates the experimental protocol. Female DBA/2 mice were injected i.p. with 10⁶ tum⁻ P911 cells and once a week with 0.5 mg MM12A1.6, C17.8, or control IgG2a AH9R3A mAb. To verify the immunogenic nature of P911 rejection in control mice, a group was irradiated with 6 Gy from a ¹³⁷Cs source. (B) Mice were monitored daily and overall survivals from two independent experiments are depicted with 11 mice for isotype group; 11 mice for MM12A1.6 group; seven mice for C17.8 group and 12 mice for irradiation control group (***p < 0.001 by the log‐rank test).
MM12A1.6 impairs rejection of an immunogenic mouse P815 tumor variant. (A) Schematic representation illustrates the experimental protocol. Female DBA/2 mice were injected i.p. with 10⁶ tum⁻ P911 cells and once a week with 0.5 mg MM12A1.6, C17.8, or control IgG2a AH9R3A mAb. To verify the immunogenic nature of P911 rejection in control mice, a group was irradiated with 6 Gy from a ¹³⁷Cs source. (B) Mice were monitored daily and overall survivals from two independent experiments are depicted with 11 mice for isotype group; 11 mice for MM12A1.6 group; seven mice for C17.8 group and 12 mice for irradiation control group (***p < 0.001 by the log‐rank test).
… 
MM12A1.6 improves skin graft survival but does not prevent parent to F1 immune aggression model. (A) Schematic representation of the skin graft protocol. Male B6 skins were grafted on female B6 mice that were treated with MM12A1.6 or control IgG2a AH9R3A mAb (0.5 mg Ab every week throughout the duration of the experiment). As a control, female skin grafts were included. (B) Graft rejection was scored daily. One representative experiment of two is shown with 10–11 mice for male skin + isotype group; nine to ten mice for male skin + MM12A1.6 group; four to seven mice for female skin group per experiment (**p < 0.01 by log‐rank test). (C) Schematic representation of the parent to F1 spleen cell transfer protocol. B6D2F1 mice were injected i.p. with 80 million B6 spleen cells and treated with MM12A1.6 or control IgG2a AH9R3A mAb (0.5 mg) i.p. just before spleen cell transfer and every 7 days subsequently. Weight loss (D) and survival (E) were monitored daily. One representative experiment of three is shown with five to seven mice for isotype group and six to eight mice for MM12A1.6 group per experiment. Statistical analysis was performed with ANOVA–Bonferroni posttest and log‐rank test and error bars of weight change graph represent mean ± SEM. (F) B6D2F1 mice were injected i.p. with 80 million B6 spleen cells and treated with MM12A1.6 or C17.8 mAb (0.5 mg) i.p. just before spleen cell transfer and every 7 days subsequently. Survival was monitored daily. One representative experiment of two is shown with five to six mice for C17.8 group and five to six mice for MM12A1.6 group per experiment. Statistical analysis was performed with log‐rank test (*p < 0.05).
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MM12A1.6 improves skin graft survival but does not prevent parent to F1 immune aggression model. (A) Schematic representation of the skin graft protocol. Male B6 skins were grafted on female B6 mice that were treated with MM12A1.6 or control IgG2a AH9R3A mAb (0.5 mg Ab every week throughout the duration of the experiment). As a control, female skin grafts were included. (B) Graft rejection was scored daily. One representative experiment of two is shown with 10–11 mice for male skin + isotype group; nine to ten mice for male skin + MM12A1.6 group; four to seven mice for female skin group per experiment (**p < 0.01 by log‐rank test). (C) Schematic representation of the parent to F1 spleen cell transfer protocol. B6D2F1 mice were injected i.p. with 80 million B6 spleen cells and treated with MM12A1.6 or control IgG2a AH9R3A mAb (0.5 mg) i.p. just before spleen cell transfer and every 7 days subsequently. Weight loss (D) and survival (E) were monitored daily. One representative experiment of three is shown with five to seven mice for isotype group and six to eight mice for MM12A1.6 group per experiment. Statistical analysis was performed with ANOVA–Bonferroni posttest and log‐rank test and error bars of weight change graph represent mean ± SEM. (F) B6D2F1 mice were injected i.p. with 80 million B6 spleen cells and treated with MM12A1.6 or C17.8 mAb (0.5 mg) i.p. just before spleen cell transfer and every 7 days subsequently. Survival was monitored daily. One representative experiment of two is shown with five to six mice for C17.8 group and five to six mice for MM12A1.6 group per experiment. Statistical analysis was performed with log‐rank test (*p < 0.05).
… 
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Basic
1482 Mélanie Gaignage et al. Eur. J. Immunol. 2021. 51: 1482–1493
DOI: 10.1002/eji.202048936
Immunomodulation and immune therapies
Research Article
Novel antibodies that selectively block mouse IL-12
enable the re-evaluation of the role of IL-12 in immune
protection and pathology
Mélanie Gaignage** 1 , Catherine Uyttenhove** 1,2 , Lindsay L. Jones3,
Christophe Bourdeaux1,PamélaChéou
1, Mohamed F. Mandour1,4 ,
Jean-Paul Coutelier1, Dario A.A. Vignali3,5,6,7 and Jacques Van
Snick1,2
1de Duve Institute, Université de Louvain, Brussels, Belgium
2Ludwig Cancer Research, Brussels, Belgium
3Department of Immunology, St. Jude Children’s Research Hospital, Memphis, TN, USA
4Department of Clinical Pathology, Faculty of Medicine, Suez Canal University, Ismailia, Egypt
5Department of Immunology, School of Medicine, University of Pittsburgh, Pittsburgh, PA,
USA
6Tumor Microenvironment Center, UPMC Hillman Cancer Center, Pittsburgh, PA, USA
7Cancer Immunology and Immunotherapy Program, UPMC Hillman Cancer Center,
Pittsburgh, PA, USA
The dimeric cytokine IL-12 is important in the control of various infections but also con-
tributes to the pathology of certain diseases making it a potential target for therapy.
However, its specic inhibition with antibodies is complicated by the fact that its two
subunits are present in other cytokines: p40 in IL-23 and p35 in IL-35. This has led to
erroneous conclusions like the alleged implication of IL-12 in experimental autoimmune
encephalomyelitis (EAE). Here, we report the development of a mouse anti-mouse IL-12
vaccine and the production of monoclonal antibodies (mAbs) that do not react with p40
or p35 (in IL-35) but specically recognize and functionally inhibit the IL-12 heterodimer.
Using one of these mAbs, MM12A1.6, that strongly inhibited IFN-γproduction and LPS-
induced septic shock after viral infection, we demonstrate the critical role played by IL-12
in the rejection of male skin graft by female C57BL/6 syngeneic recipients and in the clear-
ance of an immunogenic mastocytoma tumor variant by DBA/2 mice, but not in a parent
to F1 immune aggression model nor in MOG-induced EAE, which was clearly prevented
by anti-p40 mAb C17.8. Given this selective inhibition of IL-12, these mAbs provide new
options for reassessing IL-12 function in vivo.
Keywords: EAE rIL-12 rSepsis rTransplantation rTumor immunity
Additional supporting information may be found online in the Supporting Information section
at the end of the article.
Correspondence: Dr. Jacques Van Snick and Mélanie Gaignage
e-mail: jacques.vansnick@bru.licr.org; melanie.gaignage@uclouvain.be
**Both the authors contributed equally to this work.
© 2021 Wiley-VCH GmbH www.eji-journal.eu
... A similar pathogenic effect of IFN-g was reported after LCMV infection [18,19] and was probably related to its ability to enhance TNF production by macrophages. IL-12, which stimulates IFN-g production, notably by NK cells, therefore also plays a central role in the virally enhanced susceptibility to LPS [19,25]. Both IFN-g and TNF are involved in the overproduction of IL-1b that is required for shock induction [13]. ...
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B lymphocytes have critical roles as positive and negative regulators of immunity. Their inhibitory function has been associated primarily with interleukin 10 (IL-10) because B-cell-derived IL-10 can protect against autoimmune disease and increase susceptibility to pathogens. Here we identify IL-35-producing B cells as key players in the negative regulation of immunity. Mice in which only B cells did not express IL-35 lost their ability to recover from the T-cell-mediated demyelinating autoimmune disease experimental autoimmune encephalomyelitis (EAE). In contrast, these mice displayed a markedly improved resistance to infection with the intracellular bacterial pathogen Salmonella enterica serovar Typhimurium as shown by their superior containment of the bacterial growth and their prolonged survival after primary infection, and upon secondary challenge, compared to control mice. The increased immunity found in mice lacking IL-35 production by B cells was associated with a higher activation of macrophages and inflammatory T cells, as well as an increased function of B cells as antigen-presenting cells (APCs). During Salmonella infection, IL-35- and IL-10-producing B cells corresponded to two largely distinct sets of surface-IgM(+)CD138(hi)TACI(+)CXCR4(+)CD1d(int)Tim1(int) plasma cells expressing the transcription factor Blimp1 (also known as Prdm1). During EAE, CD138(+) plasma cells were also the main source of B-cell-derived IL-35 and IL-10. Collectively, our data show the importance of IL-35-producing B cells in regulation of immunity and highlight IL-35 production by B cells as a potential therapeutic target for autoimmune and infectious diseases. This study reveals the central role of activated B cells, particularly plasma cells, and their production of cytokines in the regulation of immune responses in health and disease.
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Local immunostimulation leading to rejection of accepted male skin grafts by female mice as a model for cancer immunotherapy
Article
Full-text available
  • Feb 2014
  • P NATL ACAD SCI USA
Significance Many cancer patients mount a spontaneous T-lymphocyte response against their tumor. In metastatic progressing patients, this response has evidently been incomplete. Moreover, it has stalled, and an immunosuppressive environment appears to be present within the tumor. This may constitute a major factor preventing success of immunotherapies such as antitumoral vaccination. Similar stalled responses are occasionnally observed in the area of tissue transplantation. In a mouse strain named CBA, this phenomenon is observed when male skin is grafted onto females. We used this system to find combinations of cytokines and Toll receptor ligands that, when applied locally, relieve the immunosuppression and reactivate the response. The combinations reported here may prove generally useful to boost the efficacy of cancer immunotherapy.
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Blockade of interleukin 27 signaling reduces GVHD in mice by augmenting Treg reconstitution and stabilizing FOXP3 expression
Article
  • Aug 2016
Key Points Blockade of IL-27 signaling mitigates the severity of GVHD by recalibrating the effector and regulatory arms of the immune system. Inhibition of IL-27 augments the reconstitution of CD4+ and CD8+ regulatory T cells and increases the stability of Foxp3 expression.
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HY peptides modulate transplantation responses to skin allografts
Article
  • Nov 2002
  • INT IMMUNOL
Injection of female C57BL/6 mice with immature female bone marrow-derived dendritic cells (BMDC) pulsed with a single immunodominant HYDb Uty peptide, WMHHNMDLI, induces prolonged survival of syngeneic male skin grafts. In contrast, injection of immature female BMDC pulsed with a single MHC class I-restricted HYAb Dby peptide, NAGFNSNRANSSRSS, causes immunization similar to that following injection of male cells. Tolerance induced by HYDb Uty peptide pretreatment is not characterized by clonal deletion: long-term tolerant mice maintain circulating HYDb Uty tetramer+ T cells which expand following exposure to male cells in vivo or in vitro. Tolerance to male skin grafts can be adoptively transferred into neonatal females with splenocytes from tolerant donors. Tolerance is specific—third-party skin grafts are rejected. We propose that tolerance in this model is initiated by cognate interaction of HYDb Uty-specific CD8+ T cells with their ligand, presented either on the injected immature BMDC or on recipient DC. This interaction leads to incomplete activation of the CD8+ T cells resulting in diminished responsiveness of CD4+ and CD8+ T cells specific for HY peptide epitopes subsequently presented on the male graft.
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Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. II. T lymphocyte-mediated cytolysis
Article
  • Nov 1980
Tumor cell variants that were rejected by syngeneic mice (tum-) were obtained from mastocytoma P815 by mutagenesis (as described in the accompanying report (13). A considerable T lymphocyte-mediated lysis was observed upon incubation of these tum- variants with peritoneal exudate cells collected a few days after an intraperitoneal challenge of immune animals. Spleen cells from these animals were cytolytic after stimulation in vitro with the immunizing variant. New antigens, absent from the original P815 tum+ cells, were detected on 15 of the 21 tum- variants that were tested. All these antigens appeared to be different. No new antigen was detected on any of 10 mutagenized P815 clones that had retained their ability to form tumors. We compared the evidence obtained in vivo and in vitro for the presence of specific antigens on five tum- variants. Three variants were shown both in vivo and in vitro to carry an individual antigen. One showed no specificity either in vivo or in vitro. However, for one variant, no specificity was observed in vivo, although cytolysis tests demonstrated the existence of a singular antigenic specificity.
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Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. I. Rejection by syngeneic mice
Article
  • Nov 1980
We have reported that it is possible to obtain variants that are incapable of forming progressive tumour in syngeneic mice (tum-) by mutagenesis and cloning of a teratocarcinoma and a Lewis lung carcinoma cell line. These observations were extended to the ascitic P815 mastocytoma of mouse strain DB/2. After a treatment with the mutagen N-methyl-N"-nitro-N-nitrosoguanidine, we obtained a high frequency (14%) of tum- clones. A colony assay indicated that after a period of rapid multiplication extending to approximately 10 d after injection, the P815 tum- cells were rejected by a process that was usually completed by day 15. No rejection was observed in sublethally irradiated animals. The immunological nature of the rejection of the P815 variants was further inferred because, upon rejection, the mice acquired a radioresistant specific protection that could be transferred adoptively with spleen cells. Cross-immunization patterns demonstrated the presence of singular antigenic specificities on three of the five variants that were examined. In addition, a common antigen was found on all the tum- variants and the original cells that were capable of forming progressive tumors in syngeneic mice (tum+). Mice injected with tum- cells were significantly protected against a tum+ challenge, even though no significant protection was generated by irradiated tum+ cells. A study of the T lymphocyte-mediated cytolysis against the P815 variants described here is presented in the accompanying report (9).
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IL-27p28 is essential for parent-to-F1 acute graft-versus-host disease
Article
  • Jul 2014
  • EUR J IMMUNOL
Acute graft versus host disease (aGVHD) remains a life-threatening complication of bone marrow transplantation. Here we show that IL-27, member of the IL-12 cytokine family, plays an essential role in a parent-to-F1 murine aGVHD model, using B6 mice as parents and B6D2 mice as F1 recipients. IL-27 is transiently detectable in the serum of B6D2 recipients of B6 spleen cells, with a peak at day 10. Treatment with anti-IL-27p28 mAb MM27.7B1 (αp28Ab), at the time of and six days after B6 cell transfer, blocked GVHD. Protection was associated with host cell survival and undiminished engraftment of donor cells, lack of host B-cell depletion, increased Th2-type immunoglobulin production, a decrease in serum IFN-γ, a drop in anti-H-2Dd cytotoxic T lymphocyte activity and an increase in Foxp3+ T cells. We therefore conclude that IL-27 plays a critical role in the parent-to-F1 model of aGVHD and that blocking IL-27 could have therapeutic relevance.This article is protected by copyright. All rights reserved
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