Application of area scaling analysis to identify natural killer cell and monocyte involvement in the GranToxiLux antibody dependent cell‐mediated cytotoxicity assay

Pollara, Justin; Orlandi, Chiara; Beck, Charles; Edwards, R. Whitney; Hu, Yi; Liu, Shuying; Wang, Shixia; Koup, Richard A.; Denny, Thomas N.; Lu, Shan; Tomaras, Georgia D.; DeVico, Anthony L.; Lewis, George K.; Ferrari, Guido

doi:10.1002/cyto.a.23348

Cited by 22 publications

(33 citation statements)

References 73 publications

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“…Using BAMA Fc arrays, we found higher levels of FcγR3a binding by immunocomplexes formed with rabbit IgG than RM IgG. Consistent with this observation, application of ASA to the results of the ADCC-GTL assay (73) demonstrated that rabbit serum preferentially recruited human NK cells. Moreover, we demonstrated that rabbit immune sera were able to induce a significantly higher frequency of NK cell CD107a degranulation (74) than RM sera.…”

Section: Discussionsupporting

confidence: 75%

“…To test this, we applied area scaling analysis (ASA) to ADCC-GTL assays performed with gp120-coated target cells (described in the legend to Fig. 4) to discriminate the ability of antibodies to recruit NK cells and monocytes present in the PBMC used as a source of effector cells (73). Using ASA, we calculated the ratio of target cells that interacted with NK cells and monocytes in the presence of serum samples.…”

Section: Resultsmentioning

confidence: 99%

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Bridging Vaccine-Induced HIV-1 Neutralizing and Effector Antibody Responses in Rabbit and Rhesus Macaque Animal Models

Pollara

Jones

Huffman

et al. 2019

J Virol

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Studies in animal models are essential prerequisites for clinical trials of candidate HIV vaccines. Small animals, such as rabbits, are used to evaluate promising strategies prior to further immunogenicity and efficacy testing in nonhuman primates. Our goal was to determine how HIV-specific vaccine-elicited antibody responses, epitope specificity, and Fc-mediated functions in the rabbit model can predict those in the rhesus macaque (RM) model. Detailed comparisons of the HIV-1-specific IgG response were performed on serum from rabbits and RM given identical modified vaccinia virus Ankara-prime/gp120-boost immunization regimens. We found that vaccine-induced neutralizing antibody, gp120-binding antibody levels and immunodominant specificities, antibody-dependent cellular phagocytosis of HIV-1 virions, and antibody-dependent cellular cytotoxicity (ADCC) responses against gp120-coated target cells were similar in rabbits and RM. However, we also identified characteristics of humoral immunity that differed across species. ADCC against HIV-infected target cells was elicited in rabbits but not in RM, and we observed differences among subdominantly targeted epitopes. Human Fc receptor binding assays and analysis of antibody-cell interactions indicated that rabbit vaccine-induced antibodies effectively recruited and activated human natural killer cells, while vaccine-elicited RM antibodies were unable to activate either human or RM NK cells. Thus, our data demonstrate that both Fc-independent and Fc-dependent functions of rabbit antibodies can be measured with commonly usedin vitroassays; however, the ability of immunogenicity studies performed in rabbits to predict responses in RM will vary depending on the particular immune parameter of interest.IMPORTANCENonneutralizing antibody functions have been associated with reduced infection risk, or control of virus replication, for HIV-1 and related viruses. It is therefore critical to evaluate development of these responses throughout all stages of preclinical testing. Rabbits are conventionally used to evaluate the ability of vaccine candidates to safely elicit antibodies that bind and neutralize HIV-1. However, it remained unexplored how effectively rabbits model the development of nonneutralizing antibody responses in primates. We administered identical HIV-1 vaccine regimens to rabbits and rhesus macaques and performed detailed comparisons of vaccine-induced antibody responses. We demonstrated that nonneutralizing HIV-specific antibody responses can be studied in the rabbit model and have identified aspects of these responses that are common, and those that are unique, to rabbits and rhesus macaques. Our findings will help determine how to best utilize preclinical rabbit and rhesus macaque models to accelerate HIV vaccine candidate testing in human trials.

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Section: Discussionsupporting

confidence: 75%

Section: Resultsmentioning

confidence: 99%

Bridging Vaccine-Induced HIV-1 Neutralizing and Effector Antibody Responses in Rabbit and Rhesus Macaque Animal Models

Pollara

Jones

Huffman

et al. 2019

J Virol

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“…In addition to ADCP, trogocytosis is another potential outcome of Fc-dependent interactions between antibodies and virus-infected cells. Two assays developed to measure HIV/SIV-specific ADCC responses have been shown to also detect antibody-mediated interactions between monocytes and virus-infected or protein-coated cells (177, 178). Both assays use fluorescent dyes to label infected or protein-coated target cells.…”

Section: Approaches To Measure Virus Adcpmentioning

confidence: 99%

Antibody-Dependent Cellular Phagocytosis in Antiviral Immune Responses

2019

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Antiviral activities of antibodies may either be dependent only on interactions between the antibody and cognate antigen, as in binding and neutralization of an infectious virion, or instead may require interactions between antibody–antigen immune complexes and immunoproteins or Fc receptor expressing immune effector cells. These Fc receptor-dependent antibody functions provide a direct link between the innate and adaptive immune systems by combining the potent antiviral activity of innate effector cells with the diversity and specificity of the adaptive humoral response. The Fc receptor-dependent function of antibody-dependent cellular phagocytosis (ADCP) provides mechanisms for clearance of virus and virus-infected cells, as well as for stimulation of downstream adaptive immune responses by facilitating antigen presentation, or by stimulating the secretion of inflammatory mediators. In this review, we discuss the properties of Fc receptors, antibodies, and effector cells that influence ADCP. We also provide and interpret evidence from studies that support a potential role for ADCP in either inhibiting or enhancing viral infection. Finally, we describe current approaches used to measure antiviral ADCP and discuss considerations for the translation of studies performed in animal models. We propose that additional investigation into the role of ADCP in protective viral responses, the specific virus epitopes targeted by ADCP antibodies, and the types of phagocytes and Fc receptors involved in ADCP at sites of virus infection will provide insight into strategies to successfully leverage this important immune response for improved antiviral immunity through rational vaccine design.

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“…The association of FcγRIIa binding with HIV-1 risk also remained significant in the IgA interaction model (OR = 0.405 [IgA -], P = 0.001; OR = 0.682 [IgA + ], P = 0.002) with the interaction P value being 0.200. Env IgG3 breadth is a weight-systemic HIV-1 infection, including clearance of virions that may decrease the number of functionally infectious virions to infect new cells and phagocytosis or trogocytosis of infected cells (54,55) that may limit their capacity to produce and spread new virions. Although our analysis showed no impact from IgG3, timing of exposure may be important to any impact on VL due to the rapid decline of Env IgG3 following vaccination (47).…”

Section: Volume 129 Number 11 November 2019mentioning

confidence: 99%