Chitinase and Fizz family members are a generalized feature of nematode infection with selective upregulation of Ym1 and Fizz1 by antigen-presenting cells - PubMed (original) (raw)

Chitinase and Fizz family members are a generalized feature of nematode infection with selective upregulation of Ym1 and Fizz1 by antigen-presenting cells

Meera G Nair et al. Infect Immun. 2005 Jan.

Abstract

Ym1 and Fizz1 are secreted proteins that have been identified in a variety of Th2-mediated inflammatory settings. We originally found Ym1 and Fizz1 as highly expressed macrophage genes in a Brugia malayi infection model. Here, we show that their expression is a generalized feature of nematode infection and that they are induced at the site of infection with both the tissue nematode Litomosoides sigmodontis and the gastrointestinal nematode Nippostrongylus brasiliensis. At the sites of infection with N. brasiliensis, we also observed induction of other chitinase and Fizz family members (ChaFFs): acidic mammalian chitinase (AMCase) and Fizz2. The high expression of both Ym1 and AMCase in the lungs of infected mice suggests that abundant chitinase production is an important feature of Th2 immune responses in the lung. In addition to expression of ChaFFs in the tissues, Ym1 and Fizz1 expression was observed in the lymph nodes. Expression both in vitro and in vivo was restricted to antigen-presenting cells, with the highest expression in B cells and macrophages. ChaFFs may therefore be important effector or wound-repair molecules at the site of nematode infection, with potential regulatory roles for Ym1 and Fizz1 in the draining lymph nodes.

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Figures

FIG. 1.

FIG. 1.

Fizz1 and Ym1 gene expression reflects protein levels. A. Western blot analysis of the peritoneal lavage fluid from individual mice. C57 or IL-4−/− mice were infected with B. malayi (imp) or injected with thioglycolate (cont). B. Time course of Fizz1 and Ym1 expression following sham surgery or B. malayi implant (Imp) of C57 mice by Western blot analysis of peritoneal lavage fluid and real-time RT-PCR of the peritoneal exudate cells. Expression is shown as a percentage of pooled B. malayi NeMφ cDNA (± standard deviations [SD] from groups of five mice). An asterisk indicates a significant difference (P < 0.05) between the implanted and sham surgery groups at the same time point.

FIG. 2.

FIG. 2.

Fizz1 and Ym1 induction during chronic infection with the filarial nematode L. sigmodontis at both the site of infection and draining LN. A, B. Real-time RT-PCR quantification of Fizz1 and Ym1 expression in thoracic lavage and draining LN cells 60 days postinfection with L. sigmodontis. Expression is shown as a percentage of pooled B. malayi NeMφ cDNA (± SD from groups of five mice). (C) ScaI restriction digest performed on the Ym PCR products from thoracic lavage (TL) cells and LN cells from infected mice (uc, uncut control; c, cut with ScaI). These data are representative of two separate experiments.

FIG. 3.

FIG. 3.

Infection with N. brasiliensis upregulates expression of Fizz and chitinases in multiple tissues. Real-time RT-PCR quantification of Fizz1 and Fizz2 (A) and Ym1 and AMCase (B) in the lung and gut tissue of naïve and BALB/c mice infected with N. brasiliensis for 6 days is shown. Expression was measured as the percentage of the highest-expressing infected tissue sample (± SD from groups of five mice). C. Sca1 restriction digest performed on the Ym PCR products of cDNA of both infected tissues. u.d., undetected by 50 amplification cycles; u.c., uncut; c., cut.

FIG. 4.

FIG. 4.

Fizz1 and Ym1 expression is induced in Th2-activated dendritic cells (A), macrophages (B), and B cells but not in T-helper cells (C). Bone marrow-derived Mφ, DC, and purified splenic B cells were left untreated (UT) or were treated with IL-4 overnight. Resting Th2 cells (rest) and Th2 cells activated with specific antigen for 3 days (act.) were obtained for expression analysis. Th1-polarized T cells were obtained by activation with immunogenic peptide over 3 weeks. Expression (mean of replicate samples) was measured by real-time RT-PCR as a percentage of pooled B. malayi NeMφ cDNA. In antigen-presenting cells, Ym1 was the sole Ym transcript observed (D). u.d., undetected by 50 amplification cycles. These data are representative of two separate experiments.

FIG. 5.

FIG. 5.

Fizz1 and Ym1 are induced in vivo in the draining lymph nodes of _B. malayi_-implanted mice. The draining lymph nodes from control mice injected with thioglycolate (cont LN) and mice implanted with B. malayi (imp LN) were recovered and prepared for gene expression and Western blot analysis. A, B. Real-time RT-PCR shows the increase in fluorescence intensity during amplification of β-actin, Fizz1, and Ym1. C. Gene expression as the percentage of pooled NeMφ cDNA (± SD from replicate samples). D. Western blot analysis for Fizz1 and Ym1 in 5 μg of protein of lymph node cell lysate, NeMφ lysate, and lavage fluid from _B. malayi_-implanted mice. Duplicate lanes represent individual mice. These data are representative of two separate experiments.

FIG. 6.

FIG. 6.

Fizz1 and Ym1 are expressed in vivo in antigen-presenting cells but not T lymphocytes. Purified cell populations from the pooled draining lymph nodes of six _B. malayi_-implanted mice were measured for Fizz1 (A) and Ym1 (B) by real-time PCR. Expression levels of each sample are shown as a percentage of the lymph node macrophages (mean of replicate samples). These data are representative of two separate experiments.

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