Bourgeoning literature verified the essential contribution of neuroinflammation in optic nerve injury, here, we aim to investigate the effect of lymphocyte deficiency on retinal ganglion cells (RGCs) survival after optic nerve crush (ONC). 48 wide type (WT) and 48 Rag1(-/-) mice were used to establish the ONC model. AAV2-hSyn1-eGFP was employed to inject into the vitreous body to transfect RGCs 4 weeks before ONC modeling, the confocal scanning laser ophthalmoscopy was utilized to visualize the RGCs in vivo. RBPMS, Iba-1 and GFAP expression were detected by immunofluorescence. The expression of retinal glial biomarkers was detected by qRT-PCR, and the protein expression of occludin and CD3 was detected by WB. Electroretinography and optomotor response were used to evaluate the visual function. Our results showed that a milder RGC loss and GCC thickness decrease were found in Rag1(-/-) mice than in WT mice after ONC in vivo and in vitro (p < 0.05). The morphologic and molecular feature analyses of retinal glial cells showed that the lack of lymphocytes significantly inhibited the number and activation level of microglia after ONC (p < 0.05). Besides, Occludin was significantly decreased and CD3 was upregulated at week 4 after ONC in WT mice compared with Rag1(-/-) mice (p < 0.01). Visual function assessment showed a better visual condition in Rag1(-/-) mice with ONC at week 4 (p < 0.05). Altogether, Rag1(-/-) mice with lymphocyte deficiency exhibit less RGC loss, milder retinal glial activation and better visual function when compared with WT mice after ONC.