A 3-year study was undertaken to examine the parity status, survival, and prevalence of West Nile virus (WNV) in overwintering populations of Culex pipiens pipiens collected from a hibernaculum located in a WNV endemic region in New York City. Nearly 6,000 females were collected from December through April. Parity rates were highest among females collected in December and January, ranging from 12.3% to 21.9%, depending on the year. In each year of the study, the proportion of parous females declined significantly during the course of the winter; the percentage of parous females found in April ranged from 0.9% to 10%. Results provide unequivocal evidence that parous Cx. p. pipiens females from this region of the northeastern US enter hibernacula in the fall in comparatively high proportions not previously recognized for this species, and while these females experience significant mortality during the winter, some survived to April to emerge in the spring. The absence of any detectible blood remnants in overwintering females reaffirms that blood feeding does not occur among diapausing females during the winter. The possibility that a portion of the diapausing population may be autogenous as a result of hybridization with sympatric belowground populations of Cx. p. pipiens “form molestus” is discussed. A single isolation of WNV was obtained in Vero cell culture from a pool of 50 females collected on January 11, 2007, representing an infection prevalence of 0.07% in the overwintering population in 2007 (n  =  1,370 mosquitoes, 33 pools). No isolations of WNV were made from mosquitoes collected in 2008 (n  =  1,870 mosquitoes, 190 pools) or 2009 (n  =  1,767 mosquitoes, 184 pools). Findings provide further evidence for local overwintering of WNV in diapausing Cx. p. pipiens, albeit at very low rates, consistent with the paucity of WNV-positive mosquitoes detected in June and early July despite the emergence of females from hibernacula in early May in this region.