Human VP8* mAbs neutralize rotavirus selectively in human intestinal epithelial cells
Ningguo Feng, … , B.V. Venkataram Prasad, Harry B. Greenberg
Ningguo Feng, … , B.V. Venkataram Prasad, Harry B. Greenberg
Published August 12, 2019
Citation Information: J Clin Invest. 2019;129(9):3839-3851. https://doi.org/10.1172/JCI128382.
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Research Article Virology

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

Abstract

We previously generated 32 rotavirus-specific (RV-specific) recombinant monoclonal antibodies (mAbs) derived from B cells isolated from human intestinal resections. Twenty-four of these mAbs were specific for the VP8* fragment of RV VP4, and most (20 of 24) were non-neutralizing when tested in the conventional MA104 cell–based assay. We reexamined the ability of these mAbs to neutralize RVs in human intestinal epithelial cells, including ileal enteroids and HT-29 cells. Most (18 of 20) of the “non-neutralizing” VP8* mAbs efficiently neutralized human RV in HT-29 cells or enteroids. Serum RV neutralization titers in adults and infants were significantly higher in HT-29 than MA104 cells and adsorption of these sera with recombinant VP8* lowered the neutralization titers in HT-29 but not MA104 cells. VP8* mAbs also protected suckling mice from diarrhea in an in vivo challenge model. X-ray crystallographic analysis of one VP8* mAb (mAb9) in complex with human RV VP8* revealed that the mAb interaction site was distinct from the human histo-blood group antigen binding site. Since MA104 cells are the most commonly used cell line to detect anti-RV neutralization activity, these findings suggest that prior vaccine and other studies of human RV neutralization responses may have underestimated the contribution of VP8* antibodies to the overall neutralization titer.

Authors

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

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Figure 2

Comparison of neutralization activity of human RV VP8*–specific mAb11 against WI61 (G9, P[8]) RV in ileal enteroids and HT-29, MA104, CV-1, and HEK293 cells.

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Comparison of neutralization activity of human RV VP8*–specific mAb11 ag...
Indicated cell types were infected with WI61 in the presence or absence of human VP8* mAb11 (100 ng/mL) for 16 hours. Cells were then fixed and stained with FITC-labeled polyclonal rabbit anti-RV antibody (green), Texas red–phalloidin (red), and DAPI (blue). Ileal enteroid 3D Matrigel culture with (A) or without (B) mAb. Ileal enteroid monolayer culture, basolateral infection with (C) or without (D) mAb. HT-29 cells with (E) or without (F) mAb. MA104 cells with (G) or without (H) mAb. CV-1 cells with (I) or without (J) mAb. HEK293 cells with (K) or without (L) mAb. Percentages of RV focus reduction by mAb11: ileal enteroid monolayer culture (97%), HT-29 cells (94%), MA104 cells (0%), CV-1 cells (11%), and HEK293 cells (0%). Original magnification, ×10.