Acute lymphoblastic leukemia (ALL) is the most common hematologic malignancy and the leading cause of childhood mortality. Single-nucleotide variants (SNVs) in key molecules of the immune system, such as Toll-like receptors (TLRs) and CD14 molecule, are associated with the development of several diseases. However, their role in ALL is unknown. A case-control study was performed with 152 ALL patients and 187 healthy individuals to investigate the role of SNVs in TLRs and the CD14 gene in ALL. In this study, TLR6 C > T rs5743810 [95% CI: 3.20, OR: 1.11–9.17, p = 0.003) and TRL9 C/T rs187084 (95% CI: 2.29, OR: 1.23–4.26, p = 0.000) seems to be a risk for development of ALL. In addition, the TLR4 A > G rs4986791 polymorphism was associated with protection from infectious comorbidities (IC 95%adj: 0.18, ORadj:0.02–1.50, p = 0.058) and the TRL1 T > G rs5743618 and TRL6 C > T rs5743810 polymorphisms with protection against death (95% IC: 0.17, OR: 0.04–0.79, p = 0.008; 95% IC: 0.48, OR: 0.24–0.94, p = 0.031, respectively). Our results show that SNVs in TLR genes may be involved in the pathogenesis of ALL and may influence clinical prognosis; however, further studies are necessary to elucidate the role of TLR1, TLR4, TLR5, TLR9 and CD14 polymorphisms in this disease.