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Mouse Invariant Monoclonal Antibody NKT14: A Novel Tool to Manipulate iNKT Cell Function In Vivo.

Scheuplein F, Lamont DJ, Poynter ME, Boyson JE, Serreze D, Lundblad LK, Mashal R, Schaub R - PLoS ONE (2015)

Bottom Line: Depletion of iNKT cells after sensitization had no effect on AHR in the conducting airways but did reduce AHR in the lung periphery.This result raises caution in the interpretation of studies that use animals that are genetically iNKT cell deficient from birth.These activating and depleting antibodies provide a novel tool to assess the therapeutic potential of iNKT cell manipulation.

View Article: PubMed Central - PubMed

Affiliation: NKT Therapeutics, Inc., Waltham, MA, United States of America.

ABSTRACT
Invariant Natural Killer T (iNKT) cells are a T cell subset expressing an invariant T Cell Receptor (TCR) that recognizes glycolipid antigens rather than peptides. The cells have both innate-like rapid cytokine release, and adaptive-like thymic positive selection. iNKT cell activation has been implicated in the pathogenesis of allergic asthma and inflammatory diseases, while reduced iNKT cell activation promotes infectious disease, cancer and certain autoimmune diseases such as Type 1 diabetes (T1D). Therapeutic means to reduce or deplete iNKT cells could treat inflammatory diseases, while approaches to promote their activation may have potential in certain infectious diseases, cancer or autoimmunity. Thus, we developed invariant TCR-specific monoclonal antibodies to better understand the role of iNKT cells in disease. We report here the first monoclonal antibodies specific for the mouse invariant TCR that by modifying the Fc construct can specifically deplete or activate iNKT cells in vivo in otherwise fully immuno-competent animals. We have used both the depleting and activating version of the antibody in the NOD model of T1D. As demonstrated previously using genetically iNKT cell deficient NOD mice, and in studies of glycolipid antigen activated iNKT cells in standard NOD mice, we found that antibody mediated depletion or activation of iNKT cells respectively accelerated and retarded T1D onset. In BALB/c mice, ovalbumin (OVA) mediated airway hyper-reactivity (AHR) was abrogated with iNKT cell depletion prior to OVA sensitization, confirming studies in knockout mice. Depletion of iNKT cells after sensitization had no effect on AHR in the conducting airways but did reduce AHR in the lung periphery. This result raises caution in the interpretation of studies that use animals that are genetically iNKT cell deficient from birth. These activating and depleting antibodies provide a novel tool to assess the therapeutic potential of iNKT cell manipulation.

No MeSH data available.


Related in: MedlinePlus

C57BL/6 mice (n = 3 per group) were injected i.v. with 50 μg NKT14m or i.p. with 2 μg αGalCer. 6 weeks after initial treatments mice were re-dosed with either NKT14m or αGalCer.Two hours after the second dose, mice were euthanized and splenocytes were analyzed by FACS for intracellular IFN-γ and IL-4. Mice were injected with αGalCer and re-challenged with αGalCer (A) or NKT14m (B) after 6 weeks. Mice were injected with NKT14m and then re-challenged with αGalCer (C) or NKT14m (D) after 6 weeks. % IFN-γ / IL-4+ iNKT cells across dosing groups (E).
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pone.0140729.g002: C57BL/6 mice (n = 3 per group) were injected i.v. with 50 μg NKT14m or i.p. with 2 μg αGalCer. 6 weeks after initial treatments mice were re-dosed with either NKT14m or αGalCer.Two hours after the second dose, mice were euthanized and splenocytes were analyzed by FACS for intracellular IFN-γ and IL-4. Mice were injected with αGalCer and re-challenged with αGalCer (A) or NKT14m (B) after 6 weeks. Mice were injected with NKT14m and then re-challenged with αGalCer (C) or NKT14m (D) after 6 weeks. % IFN-γ / IL-4+ iNKT cells across dosing groups (E).

Mentions: We have used both the depleting and activating version of the antibody in the NOD model of T1D. To determine the best regimen for our activating antibody NKT14m, we did a repeat-dosing experiment. The iNKT cell agonist αGalCer has been demonstrated to induce long term iNKT cell anergy. To assess anergy response and duration with our mAb we treated C57BL/6 mice with either αGalCer or NKT14m and then again 6 weeks later either with the same or reciprocal initially administered agent. As described in the literature, iNKT cells that have been activated with αGalCer are completely anergic to αGalCer or NKT14m after redosing at 6 weeks (Fig 2 top panel). In contrast, iNKT cells of animals that had been dosed with NKT14m were re-activated by either αGalCer or NKT14m (Fig 2 bottom panel). Based on this observation, we decided to dose every 6 weeks in the NOD T1D incidence study.


Mouse Invariant Monoclonal Antibody NKT14: A Novel Tool to Manipulate iNKT Cell Function In Vivo.

Scheuplein F, Lamont DJ, Poynter ME, Boyson JE, Serreze D, Lundblad LK, Mashal R, Schaub R - PLoS ONE (2015)

C57BL/6 mice (n = 3 per group) were injected i.v. with 50 μg NKT14m or i.p. with 2 μg αGalCer. 6 weeks after initial treatments mice were re-dosed with either NKT14m or αGalCer.Two hours after the second dose, mice were euthanized and splenocytes were analyzed by FACS for intracellular IFN-γ and IL-4. Mice were injected with αGalCer and re-challenged with αGalCer (A) or NKT14m (B) after 6 weeks. Mice were injected with NKT14m and then re-challenged with αGalCer (C) or NKT14m (D) after 6 weeks. % IFN-γ / IL-4+ iNKT cells across dosing groups (E).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608835&req=5

pone.0140729.g002: C57BL/6 mice (n = 3 per group) were injected i.v. with 50 μg NKT14m or i.p. with 2 μg αGalCer. 6 weeks after initial treatments mice were re-dosed with either NKT14m or αGalCer.Two hours after the second dose, mice were euthanized and splenocytes were analyzed by FACS for intracellular IFN-γ and IL-4. Mice were injected with αGalCer and re-challenged with αGalCer (A) or NKT14m (B) after 6 weeks. Mice were injected with NKT14m and then re-challenged with αGalCer (C) or NKT14m (D) after 6 weeks. % IFN-γ / IL-4+ iNKT cells across dosing groups (E).
Mentions: We have used both the depleting and activating version of the antibody in the NOD model of T1D. To determine the best regimen for our activating antibody NKT14m, we did a repeat-dosing experiment. The iNKT cell agonist αGalCer has been demonstrated to induce long term iNKT cell anergy. To assess anergy response and duration with our mAb we treated C57BL/6 mice with either αGalCer or NKT14m and then again 6 weeks later either with the same or reciprocal initially administered agent. As described in the literature, iNKT cells that have been activated with αGalCer are completely anergic to αGalCer or NKT14m after redosing at 6 weeks (Fig 2 top panel). In contrast, iNKT cells of animals that had been dosed with NKT14m were re-activated by either αGalCer or NKT14m (Fig 2 bottom panel). Based on this observation, we decided to dose every 6 weeks in the NOD T1D incidence study.

Bottom Line: Depletion of iNKT cells after sensitization had no effect on AHR in the conducting airways but did reduce AHR in the lung periphery.This result raises caution in the interpretation of studies that use animals that are genetically iNKT cell deficient from birth.These activating and depleting antibodies provide a novel tool to assess the therapeutic potential of iNKT cell manipulation.

View Article: PubMed Central - PubMed

Affiliation: NKT Therapeutics, Inc., Waltham, MA, United States of America.

ABSTRACT
Invariant Natural Killer T (iNKT) cells are a T cell subset expressing an invariant T Cell Receptor (TCR) that recognizes glycolipid antigens rather than peptides. The cells have both innate-like rapid cytokine release, and adaptive-like thymic positive selection. iNKT cell activation has been implicated in the pathogenesis of allergic asthma and inflammatory diseases, while reduced iNKT cell activation promotes infectious disease, cancer and certain autoimmune diseases such as Type 1 diabetes (T1D). Therapeutic means to reduce or deplete iNKT cells could treat inflammatory diseases, while approaches to promote their activation may have potential in certain infectious diseases, cancer or autoimmunity. Thus, we developed invariant TCR-specific monoclonal antibodies to better understand the role of iNKT cells in disease. We report here the first monoclonal antibodies specific for the mouse invariant TCR that by modifying the Fc construct can specifically deplete or activate iNKT cells in vivo in otherwise fully immuno-competent animals. We have used both the depleting and activating version of the antibody in the NOD model of T1D. As demonstrated previously using genetically iNKT cell deficient NOD mice, and in studies of glycolipid antigen activated iNKT cells in standard NOD mice, we found that antibody mediated depletion or activation of iNKT cells respectively accelerated and retarded T1D onset. In BALB/c mice, ovalbumin (OVA) mediated airway hyper-reactivity (AHR) was abrogated with iNKT cell depletion prior to OVA sensitization, confirming studies in knockout mice. Depletion of iNKT cells after sensitization had no effect on AHR in the conducting airways but did reduce AHR in the lung periphery. This result raises caution in the interpretation of studies that use animals that are genetically iNKT cell deficient from birth. These activating and depleting antibodies provide a novel tool to assess the therapeutic potential of iNKT cell manipulation.

No MeSH data available.


Related in: MedlinePlus