Vitamin D is crucial for immune regulation, and its deficiency is linked to multiple sclerosis (MS). The GC gene encodes Vitamin D Binding Protein (VDBP), which regulates vitamin D transport and bioavailability. This study examines the association of GC polymorphisms (rs7041, rs4588) with MS susceptibility and their impact on 25-hydroxyvitamin D [25(OH)D] levels in a Latvian cohort. This case-control study included 296 MS patients and 253 healthy controls. Genotyping of rs7041 and rs4588 was conducted using restriction fragment length polymorphism analysis and validated by Sanger sequencing. Plasma 25(OH)D levels were measured in 131 MS patients using an enzyme-linked immunosorbent assay. Statistical analysis included Hardy-Weinberg equilibrium testing, Fisher's exact test, allelic and genotypic frequency comparisons to assess MS risk, and the Kruskal-Wallis test for 25(OH)D level differences among genotypes. Our findings indicate that the rare rs7041-T and rs4588-A alleles, along with their corresponding haplotypes, exhibit a protective effect against MS (p < 0.001; OR = 0.65 for rs4588-A; p < 0.01; OR = 0.70 for rs7041-T). Conversely, the common rs7041-G and rs4588-C alleles were associated with an increased MS risk (p < 0.05). Individuals with the Gc1F/1F isotype had the highest average 25(OH)D levels (29.31 ng/mL), while Gc1S/2 carriers had the lowest (21.53 ng/mL). Our results indicate that GC polymorphisms may influence the susceptibility of Latvians to MS and vitamin D status.