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Title: MURINE MONOCLONAL ANTIBODIES AGAINST ESCHERICHIA COLI O4 LIPOPOLYSACCHARIDEAND H5 FLAGELLIN

Author
item Rivera Betancourt, Mildred
item Keen, James

Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/28/2001
Publication Date: 9/1/2001
Citation: Rivera Betancourt, M., Keen, J.E. 2001. Murine monoclonal antibodies against Escherichia coli O4 lipopolysaccharide and H5 flagellin. Journal of Clinical Microbiology. 39(9):3409-3413.

Interpretive Summary: Two murine monoclonal antibodies (MAb) (2C5-F10 and 8D1-H10) reactive with Escherichia coli O4 and H5 antigens, respectively, were generated and characterized. Enzyme immunoassays and immunoblots demonstrated that MAb 2C5-F10 reacted specifically with lipopolysaccharide O antigen of E. coli O4 isolates, while MAb 8D1-H10 reacted with E. coli strains expressing H5 flagella.

Technical Abstract: E. coli expressing O4 lipopolysaccharide (LPS) &/or H5 flagellin frequently cause extraintestinal infections in humans & domestic animals. E. coli O4 are common etiologic agents of human, canine, & feline genitourinary tract infections, manifested as pyelonephritis, cystitis, prostatitis, urosepsis, & asymptomatic bacteriuria. In particular, E. coli O4:H5 serotype is a potent human uropathogen, especially the disseminated virulent J96 clone. E. coli O4 &/or H5 antigens are also associated with enteric disease. Shiga toxin-producing E. coli (STEC) O4:H-negative, O4:H5, O4:H10, O2:H5 and O75:H5 strains have caused hemorrhagic colitis & hemolytic uremic syndrome cases, while non-STEC O4 & enteroaggregative O4 have caused pediatric diarrhea outbreaks. STEC O4:H-negative, O4:H4, O4:H21, O4:H25, & O2:H5 have been isolated from healthy cattle, while STEC O4 & non-STEC O4 have been associated with calves, pigs, & lambs with diarrhea. While the E. coli O4 & &H5 antigens are markers of strain pathogenic potential, O4-antigen moiety may itself function as a urovirulence factor. We generated & characterized 2 murine monoclonal antibodies (MAb, 2C5-F10 & 8D1-H10) reactive with E. coli O4 & O5 antigens, respectively. Enzyme immunoassays on 386 antigenically diverse gram-negative bacterial & immunoblots on representative isolates demonstrated that MAb 2C5-F10 reacted specifically with LPS O antigen of E. coli O4 isolates, while MAb 8D1-H10 reacted with E. coli strains expressing H5 flagella. These MAb have potential clinical use as immunodiagnostic or serotyping reagents, especially if used in combination with other phenotypic or genotype pathogenicity markers. They may be of particular utility for experimental or epidemiologic studies of extraintestinal E. coli O4:H5 infections.