The mannose-sensitive hemagglutinin (MSHA) is a type 4 pilus present in Vibrio cholerae O1 strains of the El Tor biotype, as well as in strains of serogroup O139. It has been shown to be a colonization antigen in animal models. The aim of this study was to investigate systemic and local antibody responses to MSHA in adult patients with cholera due to V. cholerae O1 and O139. Twenty-four of 28 (86%) patients with O1 cholera and 11 of 17 (65%) patients with O139 cholera showed significant increases in MSHA-specific immunoglobulin A (IgA) and IgM antibody-secreting cells (ASCs) 7 days after the onset of disease. However, the magnitude of the ASC response in O1 cholera patients was significantly higher than that in the O139 cholera patients in both IgA-producing (P = 0.015) and IgM-producing (P = 0.029) cells. Both groups of patients responded with antibody responses to MSHA in plasma, seroconverting with both IgA (63 to 70% of patients) and IgG (43 to 59% of patients) antibodies. Compared to the MSHA-specific antibody levels determined in healthy controls (n = 10), more than 90% of O1 and O139 cholera patients showed responses to MSHA of both the IgA and the IgG isotypes. About 70% of the patients in both groups also had antibody responses to MSHA in their feces. In summary, we demonstrated that MSHA is immunogenic, giving rise to both systemic and local antibodies in patients with cholera due to both O1 and O139 serogroups.