Development of an in vivo antibody-mediated killing (IVAK) model, a flow cytometric method to rapidly evaluate therapeutic antibodies

Guyre, Cheryl A.; Gomes, Danilo; Smith, Karen; Kaplan, Johanne; Perricone, Michael A.

doi:10.1016/j.jim.2008.01.002

Cited by 12 publications

(7 citation statements)

References 28 publications

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“…Assay of in vivo anti-FrCas E antibody cytolytic activity. The assay used was adapted from Guyre et al (26). Noninfected and FrCas E -infected splenocytes were labeled with 0.5 M (Sp CFSE low ) and 5 mM (SpFr CFSE high ) CSFE, respectively.…”

Section: Methodsmentioning

confidence: 99%

Long-Lasting Protective Antiviral Immunity Induced by Passive Immunotherapies Requires both Neutralizing and Effector Functions of the Administered Monoclonal Antibody

Nasser

Pelegrin

Michaud

et al. 2010

J Virol

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Using FrCasE retrovirus-infected newborn mice as a model system, we have shown recently that a longlasting antiviral immune response essential for healthy survival emerges after a short treatment with a neutralizing (667) IgG2a isotype monoclonal antibody (MAb). This suggested that the mobilization of adaptive immunity by administered MAbs is key for the success in the long term for the MAb-based passive immunotherapy of chronic viral infections. We have addressed here whether the anti-FrCas E protective endogenous immunity is the mere consequence of viral propagation blunting, which would simply give time to the immune system to react, and/or to actual immunomodulation by the MAb during the treatment. To this aim, we have compared viral replication, disease progression, and antiviral immune responses between different groups of infected mice: (i) mice treated with either the 667 MAb, its F(ab) 2 fragment, or an IgM (672) with epitopic specificity similar to that of 667 but displaying different effector functions, and (ii) mice receiving no treatment but infected with a low viral inoculum reproducing the initial viral expansion observed in their infected/667 MAb-treated counterparts. Our data show that the reduction of FrCas E propagation is insufficient on its own to induce protective immunity and support a direct immunomodulatory action of the 667 MAb. Interestingly, they also point to sequential actions of the administered MAb. In a first step, viral propagation is exclusively controlled by 667 neutralizing activity, and in a second one, this action is complemented by Fc␥R-bindingdependent mechanisms, which most likely combine infected cell cytolysis and the modulation of the antiviral endogenous immune response. Such complementary effects of administered MAbs must be taken into consideration for the improvement of future antiviral MAb-based immunotherapies.Although monoclonal antibodies (MAbs) principally have been considered for anticancer applications heretofore (62, 64), they now are increasingly being considered to treat severe acute and chronic viral infections (43,63,83). The best-studied antiviral MAbs are (i) pavalizumab, a humanized anti-respiratory syncytial virus (RSV) MAb approved by the FDA in 1998 for treating severe lower-respiratory-tract diseases in infants (45); (ii) several anti-human immunodeficiency virus (HIV) MAbs, which have been used in macaque preclinical infection models and in several human trials (4, 5, 19, 27-30, 32, 42, 50, 55, 57, 76-79); and (iii) a few anti-hepatitis C virus (HCV) MAbs, some of which currently are being tested in humans (9,22,40). However, other MAbs, some of them of human origin, also have been generated against other human viruses in recent years. Among them are antibodies against Ebola virus (75), West Nile virus (WNV) (48,53,54), cytomegalovirus (CMV) (11), avian and human influenza viruses (59,60,73,74), severe acute respiratory syndrome coronavirus (SARS CoV) (81), hepatitis B virus (HBV) (31, 35), Hanta virus (80, 82), and Nipah virus (80,82). These ...

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Section: Methodsmentioning

confidence: 99%

Long-Lasting Protective Antiviral Immunity Induced by Passive Immunotherapies Requires both Neutralizing and Effector Functions of the Administered Monoclonal Antibody

Nasser

Pelegrin

Michaud

et al. 2010

J Virol

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“…In vivo antibody-mediated killing (IVAK) assays 28 were performed, using fluorescent probes carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and CellTrace Far Red DDAO-SE (DDAO-SE) (Molecular Probes). Cells were labeled with 2 mM DDAO/0.1 mM CFDA-SE (control cells) or 2 mM DDAO/2 mM CFDA-SE (target cells).…”

Section: In Vivo Antibody-mediated Killing Modelmentioning

confidence: 99%

Anti-CD20 IgA can protect mice against lymphoma development: evaluation of the direct impact of IgA and cytotoxic effector recruitment on CD20 target cells

et al. 2012

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The online version of this article has a Supplementary Appendix. BackgroundWhile most antibody-based therapies use IgG because of their well-known biological properties, some functional limitations of these antibodies call for the development of derivatives with other therapeutic functions. Although less abundant than IgG in serum, IgA is the most abundantly produced Ig class in humans. Besides the specific targeting of its dimeric form to mucosal areas, IgA was shown to recruit polymorphonuclear neutrophils against certain targets more efficiently than does IgG1. Design and MethodsIn this study, we investigated the various pathways by which anti-tumor effects can be mediated by anti-CD20 IgA against lymphoma cells. ResultsWe found that polymeric human IgA was significantly more effective than human IgG1 in mediating direct killing or growth inhibition of target cells in the absence of complement. We also demonstrated that this direct killing was able to indirectly induce the classical pathway of the complement cascade although to a lesser extent than direct recruitment of complement by IgG. Recruitment of the alternative complement pathway by specific IgA was also observed. In addition to activating complement for lysis of lymphoma cell lines or primary cells from patients with lymphoma, we showed that monomeric anti-CD20 IgA can effectively protect mice against tumor development in a passive immunization strategy and we demonstrated that this protective effect may be enhanced in mice expressing the human FcαRI receptor on their neutrophils. ConclusionsWe show that anti-CD20 IgA antibodies have original therapeutic properties against lymphoma cells, with strong direct effects, ability to recruit neutrophils for cell cytotoxicity and even recruitment of complement, although largely through an indirect way.

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“…GC have been shown to synergistically enhance the induction of IL-1 beta and IL-6 [29] and the biological effects of IL-2, interferon (IFN) gamma, granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor, and oncostatin M [30]. These immune modulating effects of GC are concentration and time dependent [31; 32] and it is clear that in addition to well-known immunosuppressive effects, GC are also able to exert modulating and enhancing effects upon the immune system [33; 34; 35]. In experimental models, both suppressive and immune enhancing effects of GC have been demonstrated experimentally for inflammatory cytokine mRNA and protein production by monocytes [11; 14; 15], phagocytosis by macrophages[16], acute-phase protein gene expression by hepatocytes[36], delayed-type hypersensitivity reactions [37] and wound healing[38].…”

Section: Introductionmentioning

confidence: 99%

Glucocorticoids regulate natural killer cell function epigenetically

Eddy

Krukowski

Janusek

et al. 2014

Cellular Immunology

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Although glucocorticoids are well known for their capacity to suppress the immune response, glucocorticoids can also promote immune responsiveness. It was the purpose of this investigation to evaluate the molecular basis for this apparent dichotomous immunologic effect. Glucocorticoid treatment of natural killer cells (NK) was shown to reduce NK cell cytolytic activity by reduction of histone promoter acetylation for perforin and granzyme B, which corresponded with reduced mRNA and protein for each. In contrast, glucocorticoid treatment increased histone acetylation at regulatory regions for interferon gamma and IL-6, as well as chromatin accessibility for each. This increase in histone acetylation was associated with increased proinflammatory cytokine mRNA and protein production upon cellular stimulation. These immunologic effects were evident at the level of the individual cell and demonstrate glucocorticoids to epigenetically reduce NK cell cytolytic activity while at the same time to prime NK cells for proinflammatory cytokine production.

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Development of an in vivo antibody-mediated killing (IVAK) model, a flow cytometric method to rapidly evaluate therapeutic antibodies

Cited by 12 publications

References 28 publications

Long-Lasting Protective Antiviral Immunity Induced by Passive Immunotherapies Requires both Neutralizing and Effector Functions of the Administered Monoclonal Antibody

Long-Lasting Protective Antiviral Immunity Induced by Passive Immunotherapies Requires both Neutralizing and Effector Functions of the Administered Monoclonal Antibody

Anti-CD20 IgA can protect mice against lymphoma development: evaluation of the direct impact of IgA and cytotoxic effector recruitment on CD20 target cells

Glucocorticoids regulate natural killer cell function epigenetically

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