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OJVRTM
Online Journal of
Veterinary Research©
Volume 7:71-77, 2003. Redacted 2018.
Theileria annulata infected
cell lines induce variable cytokine gene expression.
Manuja A1*, Malhotra DV1, Sikka
VK2, Sangwan AK3, Kumar B4,
Nichani AK1**
1Department
of Veterinary Epidemiology and Preventive Medicine, 2Department of
Biotechnology, 3Department of Veterinary Parasitology and 4Central
Institute for Research on Buffaloes, Sirsa Road, Hisar-125001, Haryana, India. *Vaccine
and Infectious Disease Organization, 120 Veterinary Road, Saskatoon, SK, S7N
5E3 Canada. email: amanuja@rediffmail.com
Manuja A, Malhotra
DV, Sikka VK, Sangwan AK,
Kumar B, Nichani AK, Theileria annulata infected
cell lines induce variable cytokine gene
expression, Onl J Vet Res., 7:71-77, 2003. Bovine tropical theileriosis, an economically important haemoprotozoan disease of cattle caused by Theileria annulata, is a hindrance to crossbreeding
programmes aimed at enhancing milk production of cattle in India and other
developing countries. Control of this disease is currently achieved by using
parasite infected attenuated cell line vaccines. However, long-term culture of cell lines is labor intensive and costly. Mechanism of attenuation of schizonts in culture is also not well understood. A
correlation has been observed between the levels of proinflammatory
cytokines produced by infected cells and the pathology produced on inoculation
in susceptible animals. We analyzed the expression of
mRNA for cytokines viz. IL-1a, IL-1b, IL-6,
IL-10, IFN-γ and TNF-a from four
Indian isolates of T. annulata
from Haryana, Rajasthan and Gujarat, by reverse transcriptase polymerase chain
reaction (RT-PCR) using specific primers for each cytokine. Cytokines
IL-1α, IL-β, IL-6, IL-10 and TNF-α yielded bands of 332, 432,
655, 733 and 500 bp, respectively for all four
isolates. None of the cell lines exhibited expression for IFN-γ. To
determine intensity of amplification and compare level of cytokine expression
amongst isolates, integrated density values for each cytokine expression from
all the cell lines were calculated and divided by their respective G3PD values.
This allowed us to identify low cytokine producing cell lines over a short
period of time, which will be further exploited to produce safe, potent and
effective alternative vaccines against tropical theileriosis
KEYWORDS: Theileria annulata,
mRNA, cytokine expression, cell line immunisation.