Combined DNA-Sm14/DNA-Hsp65 vaccination reduces tissue damage during the chronic stages. A) Collagen area and B) α-SMA area in liver granulomas of mice 69 days post infection with S. mansoni in the presence or absence of immunization with DNA-Sm14 or DNA-Sm14/DNA-Hsp65. Picrosirius staining was used to measure collagen, and immunohistochemical staining was performed to detect α-SMA. Quantitative analysis corresponding to the area occupied by collagen and α-SMA fibers were determined by digital densitometry recognition and expressed as a percentage of the total area of each granuloma. The results are expressed as the mean ± standard error of a total of 25 granulomas/group ***p < 0.001 vs Control group; #p < 0.01 vs DNA-Sm14. C) The concentration of acid-soluble collagen per total protein measured in the supernatants of the liver tissue homogenates. Soluble collagen was assayed using the Sircol method, and total protein was measured using the Bradford assay. The results are expressed as the mean ± standard error of 6-8 animals/group and are representative of two experiments *p < 0.05 vs Control group 48 days; #p < 0.01 vs Control group 69 days.