Table 1.
Summary of females in excised nodule and nodules with females with PN by treatment group. The table shows the total number of nodules evaluated 18 months after treatment, summarizing the total number of alive females and number of “live” PN females in each treatment arm. I: ivermectin, D: diethylcarbamazine, A: albendazole, IDA3: IDA treatment for 3 days and PN: pleomorphic neoplasm.
Fig 1.
H&E stained cross-sections of O. volvulus females.
A: HC female with coiled embryos in the uterus. B: PN worm with neoplasm in both uterus branches. C: PN female worm with neoplasm in one uterus branch and in the pseudocoelomic cavity. The second uterus branch is filled with degenerated embryos and was not used for LCM. D: PN female with neoplasm in one uterus branch and the pseudocoelomic cavity. The second uterus branch is empty. E: PN cells in one uterus branch and in the pseudocoelomic cavity. HC: healthy control, PN: pleomorphic neoplasm, b= body wall, co= coiled embryos i = intestine, lc = lateral chord, mu= muscle, ut =uterus, scale bar= 50 μm.
Table 2.
Sample information for the analyzed paraffin-embedded O. volvulus nodules. Three different PN and three healthy control (HC) worms were selected from the IDA trial, Ghana. Four body regions were selected for laser capture microdissection: body wall (including hypodermis, lateral chord, and muscles), intestine, uterus wall and PN tissue or embryos (morulae, coiled and stretched mf). In parenthesis is the area excised from the worm for each tissue in µm2. I: ivermectin, D: diethylcarbamazine, A: albendazole and PN: pleomorphic neoplasm, *average, assessed in technical triplicates.
Fig 2.
Venn diagram representing the number of O. volvulus-derived proteins for each tissue in the PN (A) and HC (B) worms.
The overlap region between the circles shows proteins present in two or more stages. Numbers in parenthesis represents the total protein number found for the tissue/ the number of unique proteins not detected in the other sample type (i.e., detected in PN worms but not in HC worms in (A) and in HC worms but not PN worms in (B)). PN: pleomorphic neoplasm, HC: healthy control.
Fig 3.
Comparison of protein detection in tissues from PN and HC O. volvulus.
Venn diagrams represent the total number of proteins found for each tissue. In parenthesis: average tissue area cut (in µm²)/average number of spectra counts for the sample. Percentages are from the total protein PN: pleomorphic neoplasm, HC: healthy control.
Table 3.
Significantly enriched gene ontology pathways, InterPro and KEGG domains. O. volvulus-derived proteins detected in the PN tissue, only proteins that were supported by at least two unique peptides in two biological replicates were used. The top 10 enriched terms for each functional category are shown.
Fig 4.
Protein abundance correlation between PN and HC worm tissues.
The relative abundance of each detected protein in HC embryo tissue were plotted against the relative abundance in the PN tissue. Yellow dots represent proteins that are more abundant in the PN tissue. Red dots are proteins that are only highly abundant in PN and HC embryo tissue when compared to all other HC tissues proteins. PN: pleomorphic neoplasm, HC: healthy control.
Fig 5.
Immunhistolocalization of OvDig-1 (OVOC8391) in a PN female O. volvulus.
A: Cross-section of a O. volvulus female with a few stretched Mf in the vagina, negative control using pre-immune serum. B: Same section as in A but stained (red) with an antiserum raised against OvDig-1. The protein is localized in the hypodermis, connective muscle tissue and in the neoplasm. Stretched Mf are also OvDig-1 positive (arrow). C: Cross-section of a female with PN cells in both uterus branches. The outer uterus wall, the connective tissue, muscle and PN cells are OvDig-1positive (red). D: PN in the pseudocoelom cavity with strong staining for OvDig-1. Both uterus branches are empty, constricted and not labeled. E: PN in the uterus strongly labeled for OvDig-1. PN in the pseudocoelomic cavity shows a more homogenous cell type with weak labeling. HC: healthy control, PN: pleomorphic neoplasm, i = intestine, mu= muscle, ut =uterus, scale bar= 50 μm.
Fig 6.
Immunohistolocalization of OvDig-1 (OVOC8391) in a healthy female O. volvulus.
A: Non stained longitudinal section with primary oocytes in the ovary. B: Consecutive section as in A stained for OvDig-1 with positive primary oocytes (red). C: Longitudinal section of a uterus branch with stretched Mf that are strongly positive for OvDig-1. D: Cross-section of a female with different morulae stage embryos in the uterus. The outer uterus membrane is stained, some muscle cells and connective tissue. E: Cross-Section of a female with coiled (pretzel) stages in the uterus. More advanced coiled mf are OVOC8391 positive, while unmature mf are negative, like the morulae. co= coiled embryos i = intestine, lc = lateral chord, mo= morulae, mu= muscle, ov=oocytes, ut =uterus, scale bar= 20 μm.
Table 4.
A summary of Wolbachia derived proteins detected in the O. volvulus samples. Presence of Wolbachia-derived proteins in the O. volvulus samples.