Background: Ischemic stroke (IS) is a leading cause of death and disability worldwide. Since the influence of vascular stressors prompts cross-reactivity with microbial and human HSPD1 (HSP60), which results in the degradation of vascular endothelium and intimal infiltration by mononuclear cells, HSP60 is viewed as a key player in the autoimmune theory of atherosclerosis. The aim of the study: The present pilot study investigated whether the tag SNP rs11682567 HSPD1 is associated with the risk of IS. Materials and methods: DNA samples from 1176 unrelated Russian subjects (708 IS patients and 468 healthy controls) were genotyped for tagging SNP rs11682567 in the HSPD1 gene using the MassArray-4 system. A log-additive regression model was used to interpret the associations between genotypes and IS risk. Results: We discovered that smoking, a major environmental endothelial stressor, dramatically modifies the link between rs11682567 and IS, with a relationship only being seen in smokers (risk allele G; OR = 2.11, 95% CI = 1.17-3.82; P = 0.0075). The functional annotation of this SNP revealed that risk allele G of rs11682567 impacts the expression and alternative splicing of genes that regulate brain ischemia-reperfusion injury, reactive oxygen species production, toll-like receptor 4 internalization, and pro-inflammatory properties. Moreover, allele G rs11682567 creates binding sites for transcription factors jointly involved in biological processes encompass neurogenesis, monocyte differentiation, fatty acid metabolism, cell signalling, and apoptosis. Conclusion: Thus, rs11682567 SNP in the HSPD1 gene represents a novel genetic marker of ischemic stroke, whose significance for the pathogenesis of IS is mediated by smoking.