Human antibody responses to human cytomegalovirus (HCMV) envelope glycoproteins were analyzed using immunoaffinity purified glycoproteins and Western blotting. Two families of disulfide linked glycoprotein complexes, designated gC-I and gC-II, were isolated. These complexes were reduced and their individual glycoproteins separated by polyacrylamide gel electrophoresis prior to electroblotting. The reactivity of adult convalescent sera with individual glycoproteins was compared to that of sera from congenitally infected infants. All sera tested reacted with a 52,000 molecular weight glycoprotein from these complexes, but only 75% reacted with a 93,000 to 130,000 molecular weight glycoprotein from gC-I complexes. Most adult convalescent sera reacted with glycoproteins from gC-II complexes. However, 14 of 16 infant sera did not have high enough levels of gC-II antibodies to give a positive reaction with Western blotting. A longitudinal study was done with several infants and their mothers. These studies indicated a failure by the infants and their mothers to develop detectable levels of gC-II antibodies months to years after the initial infection or after repeated stimulation with HCMV due to persistent infection. The inability of these infants to develop significant levels of gC-II antibodies was not due to an inability to respond to viral glycoproteins since they had antibodies to gC-I glycoproteins. We also determined that the strains of HCMV infecting some of these infants expressed gC-II glycoproteins. Thus, the lack of response by these infants was not due to lack of expression of gC-II glycoproteins by their infecting strain.