Background: Normal immune repertoires have a fraction of cofactor-binding antibodies. Results: Heme exposure results in acquisition of reactivity to gp120 HIV-1 in 24% of antibodies in a seronegative immune repertoire; these antibodies have less mutated variable regions. Conclusion: Human immune repertoires contain high frequency of antibodies with cofactor-induced antigen specificities. Significance: This work enhances understanding of molecular features of heme-sensitive Abs and their contribution to diversification of the immune repertoires.