Eosinophils in the cerebrospinal fluid of mice infected with Angiostrongylus cantonensis are resistant to apoptosis.
Interleukin-5 (IL-5) transgenic mice were used to assess the immunological features of CSF eosinophils from mice infected with Angiostrongylus cantonensis. CSF eosinophils were hypodense by day 14 post infection (p.i.). CSF eosinophils survived longer in vitro than peritoneal eosinophils collected from cadmium sulphate (CdSO(4)) -treated normal IL-5 transgenic mice. Apoptosis was measured by Annexin V binding and the presence of a distinct laddering pattern of DNA fragmentation on agarose electrophoresis. Regardless of the presence or absence of Actinomycin D, CSF eosinophils collected from IL-5 transgenic mice from days 15-36 p.i. exhibited less apoptosis than peritoneal eosinophils collected from uninfected IL-5 transgenic mice. CSF eosinophils collected from A. cantonensis infected C57BL/6 mice at days 15-34 p.i. showed elongation of survival time and less apoptosis during in vitro cultivation. Reduced apoptosis was noted only in CSF eosinophils, but not in peritoneal eosinophils recovered from the same infected IL-5 transgenic mice. CPP32/Caspase 3 activity of cultured peritoneal eosinophils from both infected and uninfected IL-5 transgenic mice was higher than that of cultured CSF eosinophils. Stimulation with A23187 readily induced apoptosis of peritoneal eosinophils, but not CSF eosinophils or peritoneal eosinophils cultured with mouse recombinant IL-5. The latter cells were morphologically identical to hypodense eosinophils. RT-PCR analysis indicated that bcl-2 and bcl-x(L) mRNA expression was higher in CSF eosinophils compared with peritoneal eosinophils and this expression in the latter cells was upregulated after culture with mouse recombinant IL-5. These results suggest that CSF eosinophils, shifting to hypodense status through an accumulation from peripheral blood, are resistant to apoptosis. These changes may explain the long-lasting, helminthotoxic and neurotoxic actions of CSF eosinophils in A. cantonensis infection.
Sugaya
H
H
Department of Parasitology, Akita University School of Medicine, Hondo, Akita 010-8543, Japan. hirokosi@med.akita-u.ac.jp
Abe
T
T
Yoshimura
K
K
D017185:Angiostrongylus cantonensis / Q000276:immunology*; D000818:Animals; D017304:Annexin A5 / Q000737:chemistry; D017209:Apoptosis / Q000276:immunology*; D053148:Caspase 3; D020169:Caspases / Q000032:analysis; D002499:Centrifugation, Density Gradient; D053938:DNA Fragmentation / Q000276:immunology; D003609:Dactinomycin / Q000276:immunology; D004587:Electrophoresis, Agar Gel; D004804:Eosinophils / Q000166:cytology / Q000276:immunology*; D005434:Flow Cytometry; D015848:Interleukin-5 / Q000134:cerebrospinal fluid / Q000276:immunology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D019384:Nucleic Acid Synthesis Inhibitors / Q000276:immunology; D018510:RNA, Helminth / Q000737:chemistry / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017206:Strongylida Infections / Q000134:cerebrospinal fluid / Q000276:immunology*
D017304:Annexin A5; D015848:Interleukin-5; D019384:Nucleic Acid Synthesis Inhibitors; D018510:RNA, Helminth; D003609:Dactinomycin; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases