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. 2023 Apr 21;13:6577. doi: 10.1038/s41598-023-33491-7

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Migration of inflammatory monocytes to the brain and HIV-NCI development in EcoHIV infected CCL2KO compared to wildtype C57BL/6 mice. (a) 25 days after IP infection, brain immune cells were isolated and CD11b⁺LY6G^-CD45^highLY6C⁺ monocytes were measured by flow cytometry (n = 18/group). The ratio of CD11b⁺LY6G^-CD45^highLY6C⁺ cells to total brain immune cells is shown. **p < 0.01 C57BL/6 vs. CCL2KO, t-test. In another experiment, mixed-sex CCL2KO and C57BL/6 mice infected for 25 days (n = 6/condition, repeated once) were subjected to two behavioral tests to assess NCI. (b, c) results of the FC test showing mean percentages of time ± SEM spent freezing during (a) sound-shock conditioning on Day 1 and (c) auditory-cued recall on Day 2. *p < 0.05 C57BL/6xPBS vs. C57BL/6xEcoHIV, t-test. (d–f) results of the RAWM test. RM ANOVA: (d) F(3,68) = 26.275, p < 0.001/LT2, F(3,68) = 102.232, p < 0.001/LT3, F(3,68) = 46.078, p < 0.001/LT4; (e) F(3,68) = 28.144 p < 0.001/LT2, F(3,68) = 95.413, p < 0.001/LT3, F(3,68) = 97.241, p < 0.001/LT4. For RT values, see text. t-test, **p < 0.01 C57BL/6 × PBS vs. C57BL/6 × EcoHIV. (g–i) In another experiment, HIV burdens were measured at 5, 25, and 35 days after IP infection of CCL2KO and C57BL/6 mice (n = 12/group) by testing for HIV gag DNA in spleen (g), HIV gag and vif RNA in peritoneal macrophages (h), and HIV gag DNA in the brain (i). (h) t-test/day 5 p.i., **p < 0.01 C57BL/6 and CCL2KO macrophage gag RNA vs C57BL/6 and CCL2KO macrophage vif RNA; (i) t-test/days 5, 25, 35 p.i., C57BL/6 vs CCL2KO brain gag DNA, *p < 0.05 (day 5) and **p < 0.01 (day 25, 35). (j–l) mixed-sex C57BL/6 and CCL2KO mice were infected with EcoHIV by IC injection of virus and tested by RAWM 14 days p.i. (n = 6 per condition, repeated once). RM ANOVA: (j) F(3,80) = 4.035, p < 0.001/LT2, F(3,80) = 18.804, p < 0.001/LT3, F(3,80) = 45.484, p < 0.001/LT4; (k) F(3,80) = 1.776, p < 0.001/LT2, F(3,80) = 30.909, p < 0.001/LT3, F(3,80) = 26.538 , p < 0.001/LT4. For RT values, see text. t-test: *p < 0.05 and **p < 0.01 C57BL/6 × PBS vs. C57BL/6 × EcoHIV; ^#p < 0.05 and ^#p < 0.01: CCL2KO x PBS vs. CCL2KO × EcoHIV.