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Journal of General Virology (2000), 81, 2203-2213.
© 2000 Society for General Microbiology

Animal: RNA Viruses

Growth of rotaviruses in continuous human and monkey cell lines that vary in their expression of integrins

Sarah L. Londrigan1, Marilyn J. Hewish1, Melanie J. Thomsonb,1, Georgina M. Sandersc,1, Huseyin Mustafa2 and Barbara S. Coulson1,2

Department of Microbiology and Immunology, The University of Melbourne, Parkville 3052, Victoria, Australia1
Department of Gastroenterology and Clinical Nutrition, The Royal Children’s Hospital, Parkville 3052, Victoria, Australia2

Author for correspondence: Barbara Coulson. Fax +61 3 9347 1540. e-mail b.coulson{at}microbiology.unimelb.edu.au

Rotavirus replication occurs in vivo in intestinal epithelial cells. Cell lines fully permissive to rotavirus include kidney epithelial (MA104), colonic (Caco-2) and hepatic (HepG2) types. Previously, it has been shown that cellular integrins {alpha}2{beta}1, {alpha}4{beta}1 and {alpha}X{beta}2 are involved in rotavirus cell entry. As receptor usage is a major determinant of virus tropism, the levels of cell surface expression of these integrins have now been investigated by flow cytometry on cell lines of human (Caco-2, HepG2, RD, K562) and monkey (MA104, COS-7) origin in relation to cellular susceptibility to infection with monkey and human rotaviruses. Cells supporting any replication of human rotaviruses (RD, HepG2, Caco-2, COS-7 and MA104) expressed {alpha}2{beta}1 and (when tested) {alpha}X{beta}2, whereas the non-permissive K562 cells did not express {alpha}2{beta}1, {alpha}4{beta}1 or {alpha}X{beta}2. Only RD cells expressed {alpha}4{beta}1. Although SA11 grew to higher titres in RD, HepG2, Caco-2, COS-7 and MA104 cells, this virus still replicated at a low level in K562 cells. In all cell lines tested, SA11 replicated to higher titres than did human strains, consistent with the ability of SA11 to use sialic acids as alternative receptors. Levels of cell surface {alpha}2 integrin correlated with levels of rotavirus growth. The {alpha}2 integrin relative linear median fluorescence intensity on K562, RD, COS-7, MA104 and Caco-2 cells correlated linearly with the titre of SA11 produced in these cells at 20 h after infection at a multiplicity of 0·1, and the data best fitted a sigmoidal dose–response curve (r2=1·00, P=0·005). Thus, growth of rotaviruses in these cell lines correlates with their surface expression of {alpha}2{beta}1 integrin and is consistent with their expression of {alpha}X{beta}2 and {alpha}4{beta}1 integrins.




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