The cDNAs encoding the interleukin-10 and interleukin-18 of Indian water buffalo (Bubalus bubalis) were cloned and sequenced. A 537 bp IL-10 cDNA fragment and a 623 bp IL-18 cDNA fragment were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) from concanavalin A stimulated splenocytes. Sequence analysis of these cytokines revealed high level conservation at nucleic acid and protein level. Both these cytokines also showed strict conservation in the predicted secondary structure and critical amino acid residues compared to the ruminant homologues. Basal level expression of both IL-10 and IL-18 was observed in liver, lung and spleen. The expression level of IL-10 was not affected by mitogenic stimulation, whereas IL-18 was up regulated upon stimulation. The availability of these cytokine molecules will aid in the study of their role in the immunology and pathogenesis of infections in water buffalo.