Background and aim of the study: Toxoplasmosis is one of the neglected infections which has a global distribution. Effective treatment against toxoplasmosis is hard to accomplish. Accordingly, we tried to investigate the effect of using vitamin D3 alone and in combination with nitazoxanide in Toxoplasma gondii infected mice. Materials and Methods: The study was conducted on 100 Swiss albino mice which were infected with the avirulent (ME49) Toxoplasma gondii strain. Mice were equally divided into 5 groups: (GI): Infected untreated control group; (GII): Infected and treated by spiramycin; (GIII): Infected and treated by vitamin D3; (GIV): Infected and treated by nitazoxanide and (GV): Infected and received a (vitamin D3 –nitazoxanide) combination. The therapeutic impacts of drugs were assessed using; parasitological, histopathological and immunological approaches. Results: The study showed that the vitamin D3- nitazoxanide combination induced a significant reduction in the parasitic load in the brain sections of the infected mice. We have also reported a delicate balance in the overall immune response against the parasite after using the drug combination. It was noted that vitamin D3, used alone, did not significantly reduce the parasitic load despite the notable improvement in the degree of inflammatory infiltration. Conclusion: The present study revealed that the vitamin D3–nitazoxanide combination was highly potent in restricting the parasitic load. It controlled the infection without the harmful immunopathological impacts. Thus, vitamin D3 could be a valuable candidate, either as a sole agent or as an adjunct to other anti-parasitic therapies, in areas where toxoplasmosis is endemic. Background and aim of the study: Toxoplasmosis is one of the neglected infections which has a global distribution. Effective treatment against toxoplasmosis is hard to accomplish. Accordingly, we tried to investigate the effect of using vitamin D3 alone and in combination with nitazoxanide in Toxoplasma gondii infected mice. Materials and Methods: The study was conducted on 100 Swiss albino mice which were infected with the avirulent (ME49) Toxoplasma gondii strain. Mice were equally divided into 5 groups: (GI): Infected untreated control group; (GII): Infected and treated by spiramycin; (GIII): Infected and treated by vitamin D3; (GIV): Infected and treated by nitazoxanide and (GV): Infected and received a (vitamin D3 –nitazoxanide) combination. The therapeutic impacts of drugs were assessed using; parasitological, histopathological and immunological approaches. Results: The study showed that the vitamin D3- nitazoxanide combination induced a significant reduction in the parasitic load in the brain sections of the infected mice. We have also reported a delicate balance in the overall immune response against the parasite after using the drug combination. It was noted that vitamin D3, used alone, did not significantly reduce the parasitic load despite the notable improvement in the degree of inflammatory infiltration. Conclusion: The present study revealed that the vitamin D3–nitazoxanide combination was highly potent in restricting the parasitic load. It controlled the infection without the harmful immunopathological impacts. Thus, vitamin D3 could be a valuable candidate, either as a sole agent or as an adjunct to other anti-parasitic therapies, in areas where toxoplasmosis is endemic.