Secretory IgA and T cells targeting SARS-CoV-2 spike protein are transferred to the breastmilk upon mRNA vaccination

AbstractMacrophage inflammatory protein-1α (MIP-1α) and MIP-1β are distinct but highly homologous CC chemokines produced by a variety of host cells in response to various external stimuli and share affinity for CCR5. To better elucidate the role of these CC chemokines in adaptive immunity, we have characterized the affects of MIP-1α and MIP-1β on cellular and humoral immune responses. MIP-1α stimulated strong antigen (Ag)–specific serum immunoglobulin G (IgG) and IgM responses, while MIP-1β promoted lower IgG and IgM but higher serum IgA and IgE antibody (Ab) responses. MIP-1α elevated Ag-specific IgG1 and IgG2b followed by IgG2a and IgG3 subclass responses, while MIP-1β only stimulated IgG1 and IgG2b subclasses. Correspondingly, MIP-1β produced higher titers of Ag-specific mucosal secretory IgA Ab levels when compared with MIP-1α. Splenic T cells from MIP-1α– or MIP-1β–treated mice displayed higher Ag-specific Th1 (interferon-γ [IFN-γ]) as well as selective Th2 (interleukin-5 [IL-5] and IL-6) cytokine responses than did T cells from control groups. Interestingly, mucosally derived T cells from MIP-1β–treated mice displayed higher levels of IL-4 and IL-6 compared with MIP-1α–treated mice. However, MIP-1α effectively enhanced Ag-specific cell-mediated immune responses. In correlation with their selective effects on humoral and cellular immune responses, these chemokines also differentially attract CD4+ versus CD8+ T cells and modulate CD40, CD80, and CD86 expressed by B220+ cells as well as CD28, 4-1BB, and gp39 expression by CD4+ and CD8+ T cells in a dose-dependent fashion. Taken together, these studies suggest that these CC chemokines differentially enhance mucosal and serum humoral as well as cellular immune responses.

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Combination of a Sindbis-SARS-CoV-2 spike vaccine and αOX40 antibody elicits protective immunity against SARS-CoV-2 induced disease and potentiates long-term SARS-CoV-2-specific humoral and T-cell immunity

10.1101/2021.05.28.446009 ◽

2021 ◽

Author(s):

Antonella Scaglione ◽

Silvana Opp ◽

Alicia Hurtado ◽

Christine Pampeno ◽

Ziyan Lin ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Protective Immunity ◽

T Cell Response ◽

Spike Protein ◽

Effector Memory ◽

World Population ◽

Vaccine Platform ◽

Vaccine Approach

The COVID-19 pandemic caused by the coronavirus SARS-CoV-2 is a major global public threat. Currently, a worldwide effort has been mounted to generate billions of effective SARS-CoV-2 vaccine doses to immunize the world population at record speeds. However, there is still demand for alternative effective vaccines that rapidly confer long-term protection and rely upon cost-effective, easily scaled-up manufacturing. Here, we present a Sindbis alphavirus vector (SV), transiently expressing the SARS-CoV-2 spike protein (SV.Spike), combined with the OX40 immunostimulatory antibody (OX40) as a novel, highly effective vaccine approach. We show that SV.Spike plus αOX40 elicits long-lasting neutralizing antibodies and a vigorous T cell response in mice. Protein binding, immunohistochemical and cellular infection assays all show that vaccinated mice sera inhibits spike functions. Immunophenotyping, RNA Seq transcriptome profiles and metabolic analysis indicate a reprogramming of T cells in vaccinated mice. Activated T cells were found to mobilize to lung tissue. Most importantly, SV.Spike plus αOX40 provided robust immune protection against infection with authentic coronavirus in transgenic mice expressing the human ACE2 receptor (hACE2-Tg). Finally, our immunization strategy induced strong effector memory response, potentiating protective immunity against re-exposure to SARS-CoV-2 spike protein. Our results show the potential of a new Sindbis virus-based vaccine platform to counteract waning immune response that can be used as a new candidate to combat SARS-CoV-2. Given the strong T cell responses elicited, our vaccine is likely to be effective against variants that are proving challenging, as well as, serve as a platform to develop a broader spectrum pancoronavirus vaccine. Similarly, the vaccine approach is likely to be applicable to other pathogens.

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Detection of cross-reactive IgA against SARS-CoV-2 spike 1 subunit in saliva

10.1101/2021.03.29.21253174 ◽

2021 ◽

Author(s):

Keiichi Tsukinoki ◽

Tatsuo Yamamoto ◽

Keisuke Handa ◽

Mariko Iwamiya ◽

Juri Saruta ◽

...

Keyword(s):

Breast Milk ◽

Recombinant Protein ◽

University Hospital ◽

Spike Protein ◽

Secretory Iga ◽

Enzyme Linked Immunosorbent Assay ◽

Receptor Binding Domain ◽

Blood Samples ◽

Milk Samples ◽

Mucosal Surfaces

AbstractAbundant secretory IgA (sIgA) in mucus, breast milk, and saliva provides immunity that prevents infection of mucosal surfaces. sIgA in pre-pandemic breast milk samples have been reported to cross-react with SARS-CoV-2, but whether it also occurs in saliva and, if so, whether it cross-reacts with SARS-CoV-2, has remained unknown. We aimed to clarify whether sIgA in saliva cross-reacts with SARS-CoV-2 spike 1 subunit in individuals who have not been infected with this virus. The study included 137 (male, n = 101; female, n = 36; mean age, 38.7 [24–65] years) of dentists and doctors in the Kanagawa Dental University Hospital. Saliva and blood samples were analyzed by PCR and immunochromatography for IgG and IgM, respectively. We then identified patients with saliva samples that were confirmed as PCR- and IgM-negative for COVID-19. Proportions of SARS-CoV-2 cross-reactive IgA-positive individuals were determined by enzyme-linked immunosorbent assay using a biotin-labeled spike S1-mFc recombinant protein covering the receptor-binding domain of SARS-CoV-2. The proportion of SARS-CoV-2 cross-reactive IgA-positive individuals was 46.7%, and this correlated negatively with age (r = −0.218, p = 0.01). The proportion of IgA-positive individuals ≥ 50 y was significantly lower than that of patients aged ≤ 49 y (p = 0.008). sIgA was purified from the saliva of all patients, and the salivary sIgA was found to suppress the binding of SARS-CoV-2 spike protein to the ACE-2 receptor. We found SARS-CoV-2 cross-reactive sIgA in the saliva of some participants who had never been infected with the virus, suggesting that sIgA helps prevent SARS-CoV-2 infection.

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Immune responses to SARS-CoV-2 infection in hospitalized pediatric and adult patients

Science Translational Medicine ◽

10.1126/scitranslmed.abd5487 ◽

2020 ◽

Vol 12 (564) ◽

pp. eabd5487 ◽

Cited By ~ 20

Author(s):

Carl A. Pierce ◽

Paula Preston-Hurlburt ◽

Yile Dai ◽

Clare Burn Aschner ◽

Natalia Cheshenko ◽

...

Keyword(s):

T Cells ◽

Immune Responses ◽

Pediatric Patients ◽

Neutralizing Antibody ◽

Children And Youth ◽

Spike Protein ◽

Serum Concentrations ◽

Antiviral Immune Response ◽

Antibody Titers ◽

Ifn Γ

Children and youth infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have milder disease than do adults, and even among those with the recently described multisystem inflammatory syndrome, mortality is rare. The reasons for the differences in clinical manifestations are unknown but suggest that age-dependent factors may modulate the antiviral immune response. We compared cytokine, humoral, and cellular immune responses in pediatric (children and youth, age <24 years) (n = 65) and adult (n = 60) patients with coronavirus disease 2019 (COVID-19) at a metropolitan hospital system in New York City. The pediatric patients had a shorter length of stay, decreased requirement for mechanical ventilation, and lower mortality compared to adults. The serum concentrations of interleukin-17A (IL-17A) and interferon-γ (IFN-γ), but not tumor necrosis factor–α (TNF-α) or IL-6, were inversely related to age. Adults mounted a more robust T cell response to the viral spike protein compared to pediatric patients as evidenced by increased expression of CD25+ on CD4+ T cells and the frequency of IFN-γ+ CD4+ T cells. Moreover, serum neutralizing antibody titers and antibody-dependent cellular phagocytosis were higher in adults compared to pediatric patients with COVID-19. The neutralizing antibody titer correlated positively with age and negatively with IL-17A and IFN-γ serum concentrations. There were no differences in anti-spike protein antibody titers to other human coronaviruses. Together, these findings demonstrate that the poor outcome in hospitalized adults with COVID-19 compared to children may not be attributable to a failure to generate adaptive immune responses.

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Protection of the Villus Epithelial Cells of the Small Intestine from Rotavirus Infection Does Not Require Immunoglobulin A

Journal of Virology ◽

10.1128/jvi.74.9.4102-4109.2000 ◽

2000 ◽

Vol 74 (9) ◽

pp. 4102-4109 ◽

Cited By ~ 59

Author(s):

Christine M. O'Neal ◽

Gregory R. Harriman ◽

Margaret E. Conner

Keyword(s):

T Cells ◽

Normal Control ◽

Knockout Mice ◽

Immunoglobulin A ◽

Primary Immune Response ◽

Secretory Iga ◽

Compensatory Mechanisms ◽

Rotavirus Infection ◽

Serum Iga ◽

Mucosal Pathogens

ABSTRACT Immunoglobulin A (IgA) is the primary immune response induced in the intestine by rotavirus infection, but vaccination with virus-like particles induces predominantly IgG, not IgA. To definitively assess the role of IgA in protection from rotavirus infection, IgA knockout mice, which are devoid of serum and secretory IgA, were infected and then rechallenged with murine rotavirus at either 6 weeks or 10 months. Following primary rotavirus infection, IgA knockout mice cleared virus as effectively as IgA normal control mice. Rotavirus-infected IgA knockout mice produced no serum or fecal IgA but did have high levels of antirotavirus serum IgG and IgM and fecal IgG, whereas IgA normal control mice made both serum IgA and IgG and fecal IgA. Both IgA normal and IgA knockout mice were totally protected from rotavirus challenge at 42 days. Ten months following a primary infection, both IgA normal and knockout mice still had high levels of serum and fecal antirotavirus antibody and were totally protected from rotavirus challenge. To determine if compensatory mechanisms other than IgG were responsible for protection from rotavirus infection in IgA knockout mice, mice were depleted of CD4+ T cells or CD8+ T cells. No changes in the level of protection were seen in depleted mice. These data show that fecal or systemic IgA is not essential for protection from rotavirus infection and suggest that in the absence of IgA, IgG may play a significant role in protection from mucosal pathogens.

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Targeting of Secretory IgA to Peyer’s Patch Dendritic and T Cells after Transport by Intestinal M Cells

The Journal of Immunology ◽

10.4049/jimmunol.172.5.3026 ◽

2004 ◽

Vol 172 (5) ◽

pp. 3026-3033 ◽

Cited By ~ 98

Author(s):

Jacques Rey ◽

Nathalie Garin ◽

François Spertini ◽

Blaise Corthésy

Keyword(s):

T Cells ◽

Secretory Iga ◽

M Cells ◽

Peyer’S Patch ◽

Peyer's Patch

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Immunological memory to SARS-CoV-2 assessed for up to 8 months after infection

Science ◽

10.1126/science.abf4063 ◽

2021 ◽

pp. eabf4063

Author(s):

Jennifer M. Dan ◽

Jose Mateus ◽

Yu Kato ◽

Kathryn M. Hastie ◽

Esther Dawen Yu ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

B Cell ◽

Symptom Onset ◽

Post Infection ◽

Memory B Cells ◽

Spike Protein ◽

T Cell Memory ◽

Immune Memory ◽

Specific Memory

Understanding immune memory to SARS-CoV-2 is critical for improving diagnostics and vaccines, and for assessing the likely future course of the COVID-19 pandemic. We analyzed multiple compartments of circulating immune memory to SARS-CoV-2 in 254 samples from 188 COVID-19 cases, including 43 samples at ≥ 6 months post-infection. IgG to the Spike protein was relatively stable over 6+ months. Spike-specific memory B cells were more abundant at 6 months than at 1 month post symptom onset. SARS-CoV-2-specific CD4+ T cells and CD8+ T cells declined with a half-life of 3-5 months. By studying antibody, memory B cell, CD4+ T cell, and CD8+ T cell memory to SARS-CoV-2 in an integrated manner, we observed that each component of SARS-CoV-2 immune memory exhibited distinct kinetics.

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Single-blinded Study Highlighting the Differences between the Small Intestines of Neonatal and Weaned Piglets

10.21203/rs.3.rs-26152/v2 ◽

2020 ◽

Author(s):

Chen Yuan ◽

Yuxin Jin ◽

Abid Ullah Shah ◽

En Zhang ◽

penghao Zhang ◽

...

Keyword(s):

Pattern Recognition ◽

T Cells ◽

Small Intestine ◽

Mononuclear Cells ◽

Pattern Recognition Receptors ◽

Secretory Iga ◽

Secretory Cells ◽

Weaned Piglets ◽

Neonatal Piglets ◽

Small Intestines

Abstract Background: The gut is the body’s major immune structure, and the gut mucosa, which contains intraepithelial lymphocytes (IELs) and subepithelial natural immune cells, is considered the primary site for eliciting local immune responses to foreign antigens. Pigs are susceptible to intestinal infections at all life stages; however, neonates tend to be the most susceptible. This study compared the small intestine of neonatal and weaned piglets to provide a theoretical basis for preventing intestinal infectious diseases in neonatal piglets. Results: Histological analyses of weaned piglet intestines showed increased crypt depth, higher IEL count, and larger ileal Peyer’s patches compared with those of neonates. Additionally, the ileal villi of weaned piglets were longer than those of neonatal piglets. The expression of claudin-3 and occludin protein was remarkably higher in weaned piglets than in neonatal piglets. The numbers of CD3 + T cells, goblet cells, and secretory cells were also higher in the small intestine of weaned piglets than in those of neonates. The number of secretory IgA-positive cells in the jejunum was not significantly different between neonatal and weaned piglets. The gene expression of 12 pattern recognition receptors (PRRs), such as TLR1–10, MDA5, and RIG-I in the small intestines of both neonatal and weaned piglets was also examined. The mRNA expression of most pattern recognition receptors genes in the duodenum and jejunum was higher in weaners than in neonates; however, the inverse was true in the ileum. Compared with that in weaned piglets, there were significantly fewer CD3 + , CD4 + , and CD8 + T cells from peripheral blood-mononuclear cells in neonatal piglets. Conclusions: In this study, the physical and immunological components of small intestines of neonatal and weaned piglets were investigated. Our results provide preliminary data on differences in the immune mechanisms between the small intestines of 0- and 21-day-old piglets. Future studies could focus on additional developmental stages of pigs and how the differences in their small intestines affect the animal’s response to pathogens

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CD8+ T cell epitope variations suggest a potential antigen presentation deficiency for spike protein of SARS-CoV-2

10.1101/2021.01.22.427863 ◽

2021 ◽

Author(s):

Congling Qiu ◽

Chanchan Xiao ◽

Zhigang Wang ◽

Xiongfei Chen ◽

Lijuan Gao ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Immune Responses ◽

T Cell Activation ◽

Cd8 T Cells ◽

Vaccine Development ◽

Cell Epitope ◽

Cd8 T Cell ◽

Spike Protein ◽

Target Cells

AbstractCOVID-19 is caused by a newly identified coronavirus, SARS-CoV-2, and has become a pandemic around the world. The illustration of the immune responses against SARS-CoV-2 is urgently needed for understanding the pathogenesis of the disease and its vaccine development. CD8+ T cells are critical for virus clearance and induce long lasting protection in the host. Here we identified specific HLA-A2 restricted T cell epitopes in the spike protein of SARS-CoV-2. Seven epitope peptides (n-Sp1, 2, 6, 7, 11, 13, 14) were confirmed to bind with HLA-A2 and potentially be presented by antigen presenting cells to induce host immune responses. Tetramers containing these peptides could interact with specific CD8+ T cells from convalescent COVID-19 patients, and one dominant epitope (n-Sp1) was defined. In addition, these epitopes could activate and generate epitope-specific T cells in vitro, and those activated T cells showed cytotoxicity to target cells. Meanwhile, all these epitopes exhibited high frequency of variations. Among them, n-Sp1 epitope variation 5L>F significantly decreased the proportion of specific T cell activation; n-Sp1 epitope 8L>V variant showed significantly reduced binding to HLA-A2 and decreased the proportion of n-Sp1-specific CD8+ T cell, which potentially contributes to the immune escape of SAR-CoV-2.

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Nanovesicles derived from bispecific CAR-T cells targeting the spike protein of SARS-CoV-2 for treating COVID-19

Journal of Nanobiotechnology ◽

10.1186/s12951-021-01148-0 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Tianchuan Zhu ◽

Yuchen Xiao ◽

Xiaojun Meng ◽

Lantian Tang ◽

Bin Li ◽

...

Keyword(s):

T Cells ◽

Small Molecule ◽

Neutralizing Antibodies ◽

Antiviral Drugs ◽

Spike Protein ◽

Viral Resistance ◽

Single Chain ◽

Car T Cells ◽

Small Molecule Drugs ◽

Car T

Abstract Background Considering the threat of the COVID-19 pandemic, caused by SARS-CoV-2, there is an urgent need to develop effective treatments. At present, neutralizing antibodies and small-molecule drugs such as remdesivir, the most promising compound to treat this infection, have attracted considerable attention. However, some potential problems need to be concerned including viral resistance to antibody-mediated neutralization caused by selective pressure from a single antibody treatment, the unexpected antibody-dependent enhancement (ADE) effect, and the toxic effect of small-molecule drugs. Results Here, we constructed a type of programmed nanovesicle (NV) derived from bispecific CAR-T cells that express two single-chain fragment variables (scFv), named CR3022 and B38, to target SARS-CoV-2. Nanovesicles that express both CR3022 and B38 (CR3022/B38 NVs) have a stronger ability to neutralize Spike-pseudovirus infectivity than nanovesicles that express either CR3022 or B38 alone. Notably, the co-expression of CR3022 and B38, which target different epitopes of spike protein, could reduce the incidence of viral resistance. Moreover, the lack of Fc fragments on the surface of CR3022/B38 NVs could prevent ADE effects. Furthermore, the specific binding ability to SARS-CoV-2 spike protein and the drug loading capacity of CR3022/B38 NVs can facilitate targeted delivery of remdesiver to 293 T cells overexpressing spike protein. These results suggest that CR3022/B38 NVs have the potential ability to target antiviral drugs to the main site of viral infection, thereby enhancing the antiviral ability by inhibiting intracellular viral replication and reducing adverse drug reactions. Conclusions In summary, we demonstrate that nanovesicles derived from CAR-T cells targeting the spike protein of SARS-COV-2 have the ability to neutralize Spike-pseudotyped virus and target antiviral drugs. This novel therapeutic approach may help to solve the dilemma faced by neutralizing antibodies and small-molecule drugs in the treatment of COVID-19. Graphical Abstract

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