Neutrophils are the most abundant leukocyte and play a critical role in the initial response to an Ag. Recently, their ability to contribute to adaptive immunity has been highlighted. We evaluated the ability of neutrophils from blood to contribute to the adaptive immune response in a preclinical rhesus macaque SIV vaccine trial. Replication-competent adenovirus-SIV recombinants induced neutrophil activation, B cell help markers, and enhanced ability to generate reactive oxygen species. Boosting with SIV vaccines (adjuvant together with ALVAC or DNA plus envelope protein) elicited significant neutrophil responses. Serum cytokine and chemokine levels induced correlated with the frequency of neutrophil subsets expressing IL-21, myeloperoxidase, and CD64. Post-SIV infection, neutrophils exhibited dysfunction, both phenotypically and functionally. B cells from protected and infected macaques cocultured with autologous polymorphonuclear cells, consisting primarily of neutrophils, were activated, underwent class switching, and produced Abs. This B cell help was not aided by addition of IL-10 and was largely contact dependent. Numerous genes associated with inflammation, Ab production, and chemotaxis were upregulated in the cocultured B cells. We conclude that immune stimulation by vaccination or antigenic exposure imparts a greater ability of neutrophils to contribute to the adaptive immune response. Harnessing this granulocytic response has the potential to improve vaccine efficacy.

An established sex bias in HIV pathogenesis is linked to immune responses. Recently we reported a vaccine-induced sex bias: vaccinated female but not male rhesus macaques exhibited delayed SIV acquisition. This outcome was correlated with SIV Env-specific rectal IgA, rectal memory B cells, and total rectal plasma cells (PC). To uncover additional contributing factors, using samples from the same study we investigated memory B cell population dynamics in blood, bone marrow (BM) and rectal tissue during immunization and post-challenge; IgG subtypes and antibody avidity; and Breg cell frequency and function. Few sex differences were seen in Env-specific memory B cell, plasmablast or PC frequencies in the three compartments. Males had higher IgG antibody titers and avidity indices than females. However, females had elevated levels of Env-specific IgG1, IgG2, and IgG3 antibodies compared to males. gp140-specific IgG3 antibodies of females but not males were correlated with ADCC activity against gp120 targets (p = 0.026) and with antibody-dependent phagocytic activity (p = 0.010). IgG3 antibody of females but not males also correlated with decreased peak viremia (p = 0.028). Peripheral blood CD19+CD25+ Breg cells suppressed T cell proliferation compared to CD19+CD25− cells (p=0.031), and exhibited increased IL-10 mRNA expression (p=0.031). Male macaques post-vaccination (p=0.018) and post-infection (p=0.0048) exhibited higher Breg frequencies than females. Moreover, male Breg frequencies correlated with peak viremia (p=0.0071). Our data suggest that vaccinated females developed better antibody quality, contributing to better functionality. The elevated Breg frequencies in males may have facilitated SIV acquisition.

An effective human immunodeficiency virus (HIV) vaccine has yet to be developed, and defining immune correlates of protection against HIV infection is of paramount importance to inform future vaccine design. The complement system is a component of innate immunity that can directly lyse pathogens and shape adaptive immunity. To determine if complement lysis of simian immunodeficiency virus (SIV) and/or SIV-infected cells represents a protective immune correlate against SIV infection, sera from previously vaccinated and challenged rhesus macaques were analyzed for the induction of antibody-dependent complement-mediated lysis (ADCML). Importantly, the vaccine regimen, consisting of a replication-competent adenovirus type 5 host-range mutant SIV recombinant prime followed by a monomeric gp120 or oligomeric gp140 boost, resulted in overall delayed SIV acquisition only in females. Here, sera from all vaccinated animals induced ADCML of SIV and SIV-infected cells efficiently, regardless of sex. A modest correlation of SIV lysis with a reduced infection rate in males but not females, together with a reduced peak viremia in all animals boosted with gp140, suggested a potential for influencing protective efficacy. Gag-specific IgG and gp120-specific IgG and IgM correlated with SIV lysis in females, while Env-specific IgM correlated with SIV-infected cell lysis in males, indicating sex differences in vaccine-induced antibody characteristics and function. In fact, gp120/gp140-specific antibody functional correlates between antibody-dependent cellular cytotoxicity, antibody-dependent phagocytosis, and ADCML as well as the gp120-specific IgG glycan profiles and the corresponding ADCML correlations varied depending on the sex of the vaccinees. Overall, these data suggest that sex influences vaccine-induced antibody function, which should be considered in the design of globally effective HIV vaccines in the future. An HIV vaccine would thwart the spread of HIV infection and save millions of lives. Unfortunately, the immune responses conferring universal protection from HIV infection are poorly defined. The innate immune system, including the complement system, is an evolutionarily conserved, basic means of protection from infection. Complement can prevent infection by directly lysing incoming pathogens. We found that vaccination against SIV in rhesus macaques induces antibodies that are capable of directing complement lysis of SIV and SIV-infected cells in both sexes. We also found sex differences in vaccine-induced antibody species and their functions. Overall, our data suggest that sex affects vaccine-induced antibody characteristics and function and that males and females might require different immune responses to protect against HIV infection. This information could be used to generate highly effective HIV vaccines for both sexes in the future.