Human VP8* mAbs neutralize rotavirus selectively in human intestinal epithelial cells

Feng, Ningguo; Hu, Liya; Ding, Siyuan; Sanyal, Mrinmoy; Zhao, Boyang; Sankaran, Banumathi; Ramani, Sasirekha; McNeal, Monica; Yasukawa, Linda L.; Song, Yanhua; Prasad, B. V. Venkataram; Greenberg, Harry B.

doi:10.1172/jci128382

Cited by 35 publications

(42 citation statements)

References 61 publications

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“…To further characterize the potential SARS-CoV-2 route of infection, we employed the 2D monolayer system ( 22 ), with a clear separation of apical and basal compartments. We noted that the human IECs were preferentially (> 1,000 fold) infected by the virus from the apical surface compared to the basolateral side (Fig.…”

Section: Resultsmentioning

confidence: 99%

TMPRSS2 and TMPRSS4 mediate SARS-CoV-2 infection of human small intestinal enterocytes

Zang

Castro

McCune

et al. 2020

Preprint

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Both gastrointestinal symptoms and fecal shedding of SARS-CoV-2 RNA have been frequently observed in COVID-19 patients. However, whether SARS-CoV-2 replicate in the human intestine and its clinical relevance to potential fecal-oral transmission remain unclear. Here, we demonstrate productive infection of SARS-CoV-2 in ACE2+ mature enterocytes in human small intestinal enteroids. In addition to TMPRSS2, another mucosa-specific serine protease, TMPRSS4, also enhanced SARS-CoV-2 spike fusogenic activity and mediated viral entry into host cells. However, newly synthesized viruses released into the intestinal lumen were rapidly inactivated by human colonic fluids and no infectious virus was recovered from the stool specimens of COVID-19 patients. Our results highlight the intestine as a potential site of SARS-CoV-2 replication, which may contribute to local and systemic illness and overall disease progression.

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Section: Resultsmentioning

confidence: 99%

TMPRSS2 and TMPRSS4 mediate SARS-CoV-2 infection of human small intestinal enterocytes

Zang

Castro

McCune

et al. 2020

Preprint

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“…To further characterize the potential SARS-CoV-2 route of infection, we used the 2D monolayer system (27), with a clear polarity and separation of apical and basal sides that represent the luminal and lamina propria compartments, respectively. Human IECs were preferentially (>1000-fold) infected by VSV-SARS-CoV-2 from the apical surface compared with the basolateral side ( Fig.…”

Section: Sars-cov-2 Actively Replicates In Ace2 + Human Mature Enteromentioning

confidence: 99%

TMPRSS2 and TMPRSS4 promote SARS-CoV-2 infection of human small intestinal enterocytes

et al. 2020

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Gastrointestinal symptoms and fecal shedding of SARS-CoV-2 RNA are frequently observed in COVID-19. However, it is unclear whether SARS-CoV-2 replicates in the human intestine and contributes to possible fecal-oral transmission. Here, we report productive infection of SARS-CoV-2 in ACE2 + mature enterocytes in human small intestinal enteroids. Expression of two mucosa-specific serine proteases, TMPRSS2 and TMPRSS4, facilitated SARS-CoV-2 spike fusogenic activity and promoted virus entry into host cells. We also demonstrate that viruses released into the intestinal lumen were inactivated by simulated human colonic fluid, and infectious virus was not recovered from the stool specimens of patients with COVID-19. Our results highlight the intestine as a potential site of SARS-CoV-2 replication, which may contribute to local and systemic illness and overall disease progression. RESULTS SARS-CoV-2 infects human intestinal enteroidsIn the intestine, ACE2 functions as a chaperone for the sodiumdependent neutral amino acid transporter B 0 AT1 (encoded by SLC6A19) on IECs and regulates microbial homeostasis (21,22). ACE2 expression is substantially higher in the small intestine than in all the other organs, including the lung, in both humans and mice (fig. S1, A and B). Given these data, we assessed whether SARS-CoV-2 can infect IECs as a first step for understanding its implications for fecal-oral transmission. We performed single-cell RNA sequencing (scRNA-seq) to capture the global transcriptomics in all IEC subsets in the mouse small intestinal epithelium (Fig. 1A, left). ACE2 mRNA was predominantly seen in Cd26 + Epcam + Cd44 − Cd45 − mature enterocytes (Fig. 1A, right) (23, 24). In addition, bulk RNA-seq revealed that primary human ileum enteroids had substantially higher mRNA levels of all known CoV receptors, including ACE2, than the of 10 that Ace2 high cells are also positive for Cd26 and Epcam but negative for Cd44 and Cd45. (B) Human duodenum enteroids were cultured in the Transwell monolayer system using maintenance (MAINT) or differentiation (DIFF) conditions for 3 days. Monolayers were stained for ACE2 (red) and actin (phalloidin, white). Scale bars, 32 m. (C) Human duodenum enteroids in monolayer, cultured in either maintenance (MAINT) or differentiation (DIFF) conditions, were apically infected with 1.5 × 10 5 plaque-forming units (PFU) of VSV-SARS-CoV-2 [multiplicity of infection (MOI) = 0.3] for 24 hours. The expression of VSV-N was measured by RT-qPCR and normalized to that of GAPDH. p.i., post-infection. (D) Human duodenum enteroids in 3D Matrigel were cultured in maintenance (MAINT) medium or differentiation (DIFF) medium for 3 days and infected with 2.2 × 10 5 PFU of VSV-SARS-CoV-2 for 18 hours. Enteroids were stained for virus (green), actin (phalloidin, white), and nucleus (DAPI, blue). Scale bars, 50 m. (E) Same as (C) except that virus titers were measured using a TCID 50 assay instead of viral RNA levels by qPCR. (F) Same as (D) except that human ileum enteroids were used instead. Scale bars, ...

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“…Conversely, G3P [8] becomes dominant wherein RotaTeq has been used 15 . However, G2P [4] and G9P [8] were dominated in some states of Brazil 16 . In Finland, a major reduction of rotavirus gastroenteritis among hospitalized children have been noticed after the advent of Rotataq into the National Immunization Schedule 17 .…”

Section: Introductionmentioning

confidence: 96%

“…Commonly used two licensed vaccines i.e., Rotataq (monovalent), Rotarix (pentavalent) and other vaccines in China, India, and Vietnam have been implemented [11][12][13][14] . However, even after implementation of Rotarix in the national immunization schedule, G1P [8] became dominant genotype (52%) followed by G2P [4]. Conversely, G3P [8] becomes dominant wherein RotaTeq has been used 15 .…”

Section: Introductionmentioning

confidence: 99%

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Multi valent DNA vaccine against group A human rotavirus: an in-silico investigation

Dutta

2020

Preprint

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Gastroenteritis due to single rotavirus causes huge economic loss annually.Severity of rotaviral diarrhoea among children is primarily manifested by different combinations of G and P types. Rotavirus surveillance studies resulted in two ambitious globally licensed vaccine namely, Rotarix and RotaTaq and a few other.However, post-vaccination surveillance studies indicate, vaccine failure and other complications such as intussusception, environmental enteric dysfunction, etc.Herein, we design a multivalent DNA vaccine against rotavirus and tested its efficiency by using in silico tools. Two main neutralizing rotaviral antigens i.e, VP7 and VP8 were taken into account and respectively 390, 450 known sequences of different serogroup have been analyzed to obtain a consensus sequence for epitope prediction. Epitopes specific for MHC-I and -II were predicted using IEDB and chosen based on their best IC 50 value and CPR. A good binding profile with a monoclonal antibody specific for B-cell antigens is displayed by all epitopes they were found to be non-allergenic in the human host. Ethnic specificity of the epitopes is also within acceptable range except for South African and Central American populations. We use pBI-CMV1 bidirectional mammalian expression vector to design the DNA vaccine, where we stapled manually integrated epitopes for VP7 and VP8 at MCS1 and 2 respectively. In conclusion, this study provides a new set of data for a new DNA vaccine against rotavirus.

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Human VP8* mAbs neutralize rotavirus selectively in human intestinal epithelial cells

Cited by 35 publications

References 61 publications

TMPRSS2 and TMPRSS4 mediate SARS-CoV-2 infection of human small intestinal enterocytes

TMPRSS2 and TMPRSS4 mediate SARS-CoV-2 infection of human small intestinal enterocytes

TMPRSS2 and TMPRSS4 promote SARS-CoV-2 infection of human small intestinal enterocytes

Multi valent DNA vaccine against group A human rotavirus: an in-silico investigation

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