Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia

Abstract

Patterns of invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin in Caco-2 cells (without centrifugation) were compared with previously published studies of the rabbit ileal invasion assay (RIIA) and (where relevant) a HEp-2 cell invasion assay. Optimal conditions for the use of Caco-2 cell monolayers in bacterial invasion assays were de®ned. Centrifuge-assisted attachment of bacteria to cells was not used routinely as this increased the invasiveness of known hypo-invasive strains and detachment of Caco-2 cells. Inocula with too high bacterial numbers resulted in rapid acidi®cation of media and detachment of the monolayers. The invasiveness of Typhimurium strains TML, WAKE, WII8, LT7, SL1027 and M206 in Caco-2 cells re¯ected that seen in the RIIA. The invasiveness of Choleraesuis strain A50 was similar to that in the RIIA except that bacteria grown at 378C and used without storage at 48C were slightly more invasive than those grown at 378C and stored at 48C before use. Dublin strain 3246 showed no apparent temperature-regulated invasiveness in Caco-2 cells, in contrast to the results observed in the RIIA. Dublin strain 3246 did not cleave tight junctions in the Caco-2 cell monolayer as it did in rabbit ileal epithelia both in vitro and in vivo. Three Tn phoA insertion LPS mutants of Typhimurium TML were uniformly hypo-invasive in both Caco-2 cells and the RIIA; in contrast, they were differentially invasive in HEp-2 cells. Three smooth Tn phoA insertion mutants of Typhimurium TML (invH, invG and pagC) were hypo-invasive in both the Caco-2 and HEp-2 cell invasion assays but not in the RIIA.

Identification of genetic and phenotypic differences associated with prevalent and non-prevalent Salmonella Enteritidis phage types: analysis of variation in amino acid transport

“…ml 21 . Duplicate plates of 10-day-old Caco-2 human intestinal epithelial cells (well differentiated) were set up for association assays with bacterial cultures (Bolton et al, 2000). Adhesion and invasion assays were performed in four replicates per strain and repeated at least two times.…”

Differential Phenotypic Diversity among Epidemic-Spanning Salmonella enterica Serovar Enteritidis Isolates from Humans or Animals

“…To study the interaction of S. Enteritidis isolates and human intestinal epithelium, we used the enterocyte-like Caco-2 cell line cocultured with bacteria (7,12,16,28). Motility is an important factor influencing invasion (30,40,46,47); thus, we first tested the motility of the strains in soft agar (Fig.…”

Molecular Mechanisms of Salmonella Pathogenicity: Review

“…ve ti fo id ol ma yan Sal mo nel la se ro tip le ri gibi) ba ğır sak epi te li ni iş gal eder ler ve ba ğır sak ta yo ğun nöt ro fil akı şı na ne den olur lar. 13,14 S. typhi mu ri um ta ra fın dan in dük le nen gas troen te ri tin me ka niz ma sı üze rin de yü rü tü len son ça-lış ma lar da, nöt ro fil sız ma sı (akı şı) ile ka rak te ri ze edi len yan gı lı di ya re nin bak te ri yel iş gal ve onu takip eden do ğal ba ğı şık lık ya nı tın dan ile ri gel di ği be lir len miş tir. En fek si yon es na sın da S. typhi mu rium, Sal mo nel la pa to je ni te ada sı 1 (SPI1)'de kod lanan tip III sal gı sis te mi (T3SS-1) ara cı lı ğı ile efektör pro te in le ri ni en jek te ede rek, en te ro sit le re ve mikr fold (M) hüc re le ri ne gi rer.…”

“…12,23 S. typhi mu ri um ile in san hüc re le ri ara sın da ki in te rak si yon, mak ro faj ben ze ri hüc re ler (THP-1 ve ya U937 hüc re le ri), mak ro faj lar dan tü re yen mono sit hüc re le ri ve ya bi rin cil nöt ro fil le rin, söz konu su bak te ri ile en fek te edil me si so nu cu mo del len miş tir. 13,14 Son za man da ise in san vü cudun dan alı nan do ku ör nek le ri S. typhi mu ri um ile in san ba ğır sak mu ko za sı ara sın da ki iliş ki nin ta nım-la ma sı ama cı ile kul la nıl mak ta dır. İnsan epi tel hüc-re le ri nin kul la nıl dı ğı bu ça lış ma lar da; SopB ta ra fın dan üre ti len fos fa ti di li no zi tol fos fa taz lar ile So pE-So pE2 ta ra fın dan üre ti len RhoGT Paz' la rın be ra ber ça lı şa rak S. typhi mu ri um iş ga li için ge rekli olan WASp/Scar pro te in le ri ni ak ti ve et ti ği saptan mış tır.…”