Sains Malaysiana 51(10)(2022): 3481-3493

http://doi.org/10.17576/jsm-2022-5110-29

Pencirian Jangkitan Plasmodium berghei NK65 pada Mencit ICR sebagai Model Jangkitan Malaria Teruk

(Characterisation of Plasmodium berghei NK65 Infection in ICR Mice as a Severe Malarial Infection Model)

AMATUL HAMIZAH ALI¹, WAN ROZIANOOR MOHD HASSAN², DHIANA EFANI DAHARI³, NOOR EMBI³, HASIDAH MOHD SIDEK³, RUSLIZA BASIR⁴, HANI KARTINI AGUSTAR⁵ & JALIFAH LATIP^1,*

¹Jabatan Sains Kimia, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

²Fakulti Sains Gunaan, Universiti Teknologi MARA, 40450 Shah Alam, Selangor Darul Ehsan, Malaysia

³Jabatan Sains Biologi dan Bioteknologi, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

⁴Unit Farmakologi, Jabatan Anatomi Manusia, Fakulti Perubatan dan Sains Kesihatan, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

⁵Jabatan Sains Bumi dan Alam Sekitar, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

Diserahkan: 13 Mac 2022/Diterima: 4 Julai 2022

Abstrak

Malaria teruk atau ‘severe’ kebiasaannya disebabkan oleh jangkitan Plasmodium falciparum. Jangkitan Plasmodium falciparum pada manusia boleh menyebabkan kerosakan organ, anemia teruk, komplikasi serius, koma dan kematian. Bagi tujuan memahami patogenesis malaria teruk, model haiwan digunakan dalam kajian kali ini bagi mengenal pasti sama ada gabungan hos-parasit daripada mencit ICR dengan Plasmodium berghei NK65 boleh menyebabkan jangkitan malaria teruk pada hos. Pencirian jangkitan P. berghei ANKA pernah dilakukan sebelum ini terhadap mencit ICR; walau bagaimanapun, pencirian jangkitan P. berghei NK65 secara terperinci terhadap mencit ICR dalam kajian ini adalah pertama kali dilaporkan. Inokulasi sel darah merah (RBC) terjangkit-P. berghei NK65 (2 ´ 10⁷ parasit RBC (pRBC)/mL) dilakukan terhadap mencit ICR dengan suntikan secara intraperitoneum. Pemantauan perubahan ciri fizikal seperti berat, suhu mencit, kematian mencit, pos mortem, histologi dan aras sitokin inflamasi yang terhasil selepas jangkitan direkod untuk analisis. Strain P. berghei NK65 menghasilkan jangkitan tahap teruk terhadap mencit ICR iaitu paras parasitemia melebihi 50% pada hari ke-10 selepas jangkitan diikuti kematian. Analisis histopatologi menunjukkan jangkitan ini menyebabkan perubahan pada tisu serebrum, perlekatan leukosit pada endotelium dan pensekuesteran pRBC dalam salur darah serebrum serta pendarahan intravaskular. Selepas jangkitan, pensekuesteran pRBC dan pengumpulan pigmen malaria turut dilihat pada organ utama mencit. Tambahan lagi, edema pulmonari, pembentukan membran hialin pada peparu dan pendarahan kortikal pada ginjal dilihat pada mencit terjangkit. Sitokin proinflamasi (TNF-α, IFN-γ, dan IL-18) dan sitokin antiinflamasi (IL-10 dan IL-4) juga meningkat dalam serum mencit terjangkit. Secara rumusannya, model jangkitan mencit ICR-P. berghei NK65 yang digunakan dalam kajian ini menunjukkan ciri-ciri jangkitan malaria teruk. Hasil daripada kajian ini boleh digunakan sebagai asas untuk memahami patogenesis bagi malaria teruk pada manusia dan model jangkitan malaria haiwan pada masa akan datang.

Kata kunci: Histopatologi; malaria; mencit ICR; model jangkitan haiwan; Plasmodium berghei NK65; sitokin inflamasi

Abstract

Severe malaria is commonly caused by Plasmodium falciparum infection. Plasmodium falciparum infection in human can cause organ damage, severe anaemia, serious complications, coma and death. For the purpose of understanding the pathogenesis of severe malaria, an animal model was used in this study to examine whether the combination of host-parasite from ICR mice with Plasmodium berghei NK65 caused severe malaria infection in the host. Characterisation of P. berghei ANKA infection has been performed previously on ICR mice, however, the detailed histopathological view of P. berghei NK65 infection on ICR mice in this study was first reported. Inoculation of the P. berghei NK65-infected red blood cells (RBCs) (2×10⁷ parasitised RBCs (pRBC)/mL) were performed on ICR mice by intraperitoneal injection. Changes in physical characteristics such as body weight, temperature, mortality, post-mortem, histopathology and levels of inflammatory cytokines resulting after infection were recorded for analysis. The P. berghei NK65 strain produced a severe level of infection in ICR mice i.e., the degree of parasitaemia exceeded 50% on day-10 after infection followed by death. Histopathological analysis showed that the infection caused changes in cerebral tissue, accumulation of leukocytes to the endothelium and sequestration of pRBCs in the cerebral blood vessels as well as intravascular haemorrhage. After infection, pRBC sequestration and accumulation of malaria pigments were also observed in the major organs. In addition, pulmonary oedema, hyaline membrane formation in the lungs and cortical haemorrhage in the kidneys were seen in infected mice. Proinflammatory cytokines (TNF-α, IFN-γ, and IL-18) and anti-inflammatory cytokines (IL-10 and IL-4) were also increased in the serum of infected mice. In summary, the ICR mice-P. berghei NK65 infection model used in this study showed characteristics of severe malaria infection in human. The insights from this study can be used as a basis for understanding the pathogenesis of severe malaria in human and models of rodent malarial infection in the future.

Keywords: Animal infection model; histopathology; ICR mice; inflammatory cytokines; malaria; Plasmodium berghei NK65

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^*Pengarang untuk surat-menyurat; email: jalifah@ukm.edu.my