title: Induction of TNF-alfa and CXCL-2 mRNAs in different organs of mice infected with pathogenic Leptospira.
abstract: The role of innate immune response in protection against leptospirosis is poorly understood. We examined the expression of the chemokine CXCL2/MIP-2 and the cytokine TNF-α in experimental resistant and susceptible mice models, C3H/HeJ, C3H/HePas and BALB/c strains, using a virulent strain of Leptospira interrogans serovar Copenhageni. Animals were infected intraperitoneally with 10(7) cells and the development of the disease was followed. Mortality of C3H/HeJ mice was observed whereas C3H/HePas presented jaundice and BALB/c mice remained asymptomatic. The infection was confirmed by the presence of leptospiral DNA in the organs of the animals, demonstrated by PCR. Sections of the organs were analyzed, after H&E stain. The relative expression of mRNA of chemokine CXCL2/MIP-2 and cytokine TNF-α was measured in lung, kidney and liver of the mice by qPCR. The concentrations of these proteins were measured in extracts of tissues and in serum of the animals, by ELISA. Increasing levels of transcripts and protein CXCL2/MIP-2 were detected since the first day of infection. The highest expression was observed at third day of infection in kidney, liver and lung of BALB/c mice. In C3H/HeJ the expression of CXCL2/MIP-2 was delayed, showing highest protein concentration in lung and kidney at the 5th day. Increasing in TNF-α transcripts were detected after infection, in kidney and liver of animals from the three mice strains. The expression of TNF-α protein in C3H/HeJ was also delayed, being detected in kidney and lung. Our data demonstrated that Leptospira infection stimulates early expression of CXCL2/MIP-2 and TNF-α in the resistant strain of mice. Histological analysis suggests that the expression of those molecules may be related to the influx of distinct immune cells and plays a role in the naturally acquired protective immunity.
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, Brazil. josilvab@butantan.gov.br
authors.forename: Josefa B
authors.initials: JB
authors.lastname: da Silva
authors.forename: Enéas
authors.initials: E
authors.lastname: Carvalho
authors.forename: Ambart E
authors.initials: AE
authors.lastname: Covarrubias
authors.forename: Ana Tung C
authors.initials: AT
authors.lastname: Ching
authors.forename: Vania G M
authors.initials: VG
authors.lastname: Mattaraia
authors.forename: Delhi
authors.initials: D
authors.lastname: Paiva
authors.forename: Marcelo
authors.initials: M
authors.lastname: de Franco
authors.forename: Regiane Degan
authors.initials: RD
authors.lastname: Fávaro
authors.forename: Martha M
authors.initials: MM
authors.lastname: Pereira
authors.forename: Silvio
authors.initials: S
authors.lastname: Vasconcellos
authors.forename: Telma T M
authors.initials: TT
authors.lastname: Zorn
authors.forename: Paulo Lee
authors.initials: PL
authors.lastname: Ho
authors.forename: Elizabeth A L
authors.initials: EA
authors.lastname: Martins
mesh_terms: D000818:Animals; D054426:Chemokine CXCL2 / Q000235:genetics* / Q000276:immunology; D060467:Disease Resistance; D006801:Humans; D007113:Immunity, Innate; D007668:Kidney / Q000276:immunology; D007919:Leptospira / Q000276:immunology / Q000502:physiology*; D007922:Leptospirosis / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D008099:Liver / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000276:immunology; D015854:Up-Regulation
chemical_list: D054426:Chemokine CXCL2; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha