title: Phenylhydrazine administration accelerates the development of experimental cerebral malaria.
abstract: Phenylhydrazine (PHZ) treatment is generally used to enhance parasitemia in infected mice models. Transient reticulocytosis is commonly observed in iron-deficient anemic hosts after treatment with iron supplementation, and is also associated with short-term hemolysis caused by PHZ treatment. In this study, we investigated the relationship between reticulocytosis and cerebral malaria (CM) in a murine model induced by PHZ administration before Plasmodium berghei ANKA (PbA) infection. Mortality and parasitemia were checked daily. Pro-inflammatory cytokines and IL-10 were quantified by ELISA. The expression of CXCL9, CXCL10, CCL5, and CXCR3 mRNAs was determined by real-time PCR. Brain sequestration of CD4(+) and CD8(+) T cells and populations of splenic Th1 CD4(+) T cells, dendritic cells (DCs), CD11b(+) Gr1(+) cells, and regulatory T cells (Tregs) were assessed by FACS. PHZ administration dramatically increased parasitemia from day 3 to day 5 post infection (p.i.) compared with the untreated control infected mice group; also, CM developed at day 5 p.i., compared with day 7 p.i. in untreated control infected mice, as well as significantly decreased blood-brain barrier function (P < 0.001). PHZ administration during PbA infection significantly increased the expression of CXCL9 (P <0.05) and VCAM-1 (P <0.001) in the brain, increased the expression of CXCL10, CCL5 and CXCR3, and significantly increased the recruitment of CD4(+) and CD8(+) T cells (P <0.001 and P <0.01, respectively) as well as CD11b(+) Gr1(+) cells to the brain. In addition, PHZ administration significantly increased the numbers of IL-12-secreting DCs at days 3 and 5 p.i. compared to those of untreated control infected mice (P <0.001 and P <0.01, respectively). Consequently, the activation of CD4(+) T cells, especially the expansion of the Th1 subset (P <0.05), was significantly and dramatically enhanced and was accompanied by marked increases in the production of protein and/or mRNA of the Th1-type pro-inflammatory mediators, IFN-γ and TNF-α (P <0.01 for both for protein; P <0.05 for TNF-α mRNA). Our results suggest that, compared to healthy individuals, people suffering from reticulocytosis may be more susceptible to severe malaria infection in malaria endemic areas. This has implications for the most appropriate selection of treatment, which may also cause reticulocytosis in patients living in such areas.
authors.affiliation: Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, 110001, China.
authors.forename: Xiaotong
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, 110001, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, 110001, China.
authors.forename: Yonghui
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, 110001, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Pang
authors.affiliation: Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, 110001, China.
authors.forename: Zanmei
authors.initials: Z
authors.lastname: Qi
authors.affiliation: Department of Laboratory Medicine, the First Hospital of China Medical University, Shenyang, Liaoning, China; The Key Laboratory of AIDS Immunology of Ministry of Health, the First Hospitol of China Medical University, Shenyang, Liaoning, China.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Department of Laboratory Medicine, the First Hospital of China Medical University, Shenyang, Liaoning, China; The Key Laboratory of AIDS Immunology of Ministry of Health, the First Hospitol of China Medical University, Shenyang, Liaoning, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Shang
authors.affiliation: Department of Immunology, College of Basic Medical Science, China Medical University, Shenyang, 110001, China. Electronic address: ymcao@mail.cmu.edu.cn.
authors.forename: Yaming
authors.initials: Y
authors.lastname: Cao
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000378:metabolism; D004906:Erythrocyte Count; D004909:Erythrocyte Indices; D005260:Female; D006454:Hemoglobins / Q000032:analysis; D007274:Injections, Intraperitoneal; D016779:Malaria, Cerebral / Q000139:chemically induced*; D051379:Mice; D008810:Mice, Inbred C57BL; D016877:Oxidants / Q000009:adverse effects*; D018512:Parasitemia / Q000139:chemically induced*; D010659:Phenylhydrazines / Q000009:adverse effects*; D010962:Plasmodium berghei / Q000187:drug effects* / Q000472:pathogenicity; D011897:Random Allocation; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism; D054367:Receptors, CXCR3 / Q000235:genetics / Q000378:metabolism; D012156:Reticulocytes / Q000166:cytology / Q000187:drug effects; D045262:Reticulocytosis / Q000187:drug effects* / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000187:drug effects; D015854:Up-Regulation
chemical_list: C000592874:CCR5 protein, mouse; C515188:Cxcr3 protein, mouse; D006454:Hemoglobins; D016877:Oxidants; D010659:Phenylhydrazines; D019713:Receptors, CCR5; D054367:Receptors, CXCR3; C030299:phenylhydrazine
keywords: CD11b(+) Gr1(+); Cerebral malaria; Phenylhydrazine; Plasmodium berghei; Reticulocytosis