Induction of alpha and beta chemokines by intestinal epithelial cells stimulated with Campylobacter jejuni.
OBJECTIVE
To investigate the production of dynamic alpha and beta chemokines represented by interleukin-8 (IL-8) as alpha chemokine and CCL2 (monocyte-chemoattractant protein-1, CCR2 ligand), CCL4 (macrophage-inflammatory protein-1beta, CCR5 ligand), CCL3 (macrophage-inflammatory protein-1alpha, CCR1/5 ligand), (CCL5, regulated upon activation, normal T-cell expressed and secreted (RANTES, CCR5 ligand) as beta chemokines by the human intestinal cell line INT407 stimulated with factors produced by living Campylobacter jejuni (C. jejuni) and those present within sonicated and filtrated bacteria.


METHODS
We used immunohistochemical technique modified to detect intracellular production of cytokines protein and RT-PCR to read RNA messages for evaluation of de novo cytokine synthesis.


RESULTS
Living bacteria induced increased numbers of IL-8, CCL4 and CCL2 but not CCL3 or CCL5 producing cells. Low numbers of IL-8, CCL4 and CCL2 producing cells were detected with filtrated supernatant compared to living and sonicated bacteria. A non-significant low number of chemokine producing cells was noted when comparing numbers of chemokine producing cells stimulated with living C. jejuni to those stimulated with sonicated bacteria, indicating that the triggering factors involved in stimulation with living bacteria were still active after sonication, but they were largely lost upon filtration. The mRNA signals for IL-8 were noted in conformity with its protein levels as increased IL-8 mRNA signals were registered after stimulation with living and sonicated bacteria but not with filtrated supernatant.


CONCLUSIONS
Preferential production of chemokines probably induced by membrane associate factors of C. jejuni acting on intestinal epithelial cells is presented. These chemokines are suggested to be part of an inflammatory network affecting cell types that contribute to initiation and/or resolution of the infection.
Bakhiet
Moiz
M
Department of Microbiology, Immunology and Infectious Diseases, Arabian Gulf University, Manama, Bahrain. moiz.bakhiet@medhs.ki.se
Al-Salloom
Fajer Subah
FS
Qareiballa
Ahmed
A
Bindayna
Khalid
K
Farid
Iman
I
Botta
Giuseppe A
GA
D000704:Analysis of Variance; D016123:Campylobacter jejuni / Q000276:immunology / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis*; D006801:Humans; D007124:Immunoenzyme Techniques; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018925:Chemokines; D012333:RNA, Messenger