Phosphorylating enzymes (PEs) are responsible for activating nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs) such as tenofovir (TFV) and are critical for their conversion to obtain intracellular antiviral activity. However, there are limited data available regarding the expression of PEs and their activity in the female genital tract. This work compared the messenger RNA (mRNA) expression levels of PEs in human female genital tissue, immune cells, and animal models that are commonly used in human immunodeficiency virus (HIV) research. Furthermore, the effect of contraceptive hormones and proinflammatory cytokines on tenofovir diphosphate (TFV-DP) formation and efficacy in human vaginal, epithelial, and immune cells was also evaluated. We found that human vaginal and ectocervical tissues had similar mRNA expression for seven PEs tested. Polymerase chain reaction results revealed that creatine kinase brain (CKB), mitochondrial creatine kinase 1 (CKMT1), mitochondrial creatine kinase 2 (CKMT2), adenylate kinase AK3L1 (AK4), and nucleoside diphosphate kinase 1 (NME1) exhibited a 10- to 10,000-fold higher expression level in a vaginal epithelial cell line, VK2, compared with CD4⁺ T cells (p < .05). Medroxyprogesterone acetate (MPA)/progesterone (P4) and IL-1β/IL-8 treatment resulted in altered TFV-DP levels in VK2 and PM1 cells. MPA and P4 at concentrations above 0.1 μM, as well as IL-1β and IL-8 at concentrations above 10 ng/mL, significantly decreased HIV-1_BaL inhibition in PM1 cells when 1 μM TFV was added. However, this observed effect of hormones and cytokines was abrogated when TFV concentration was raised to 1 mM. These in vitro results elucidate the role of PEs in TFV metabolism and provide information regarding differences in PE tissue expression for animal models commonly used in HIV testing. This information can be applied to better understand and interpret data obtained using these models.

中文翻译：

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与 HIV-1 性传播相关的组织和细胞中磷酸化酶的表达、活性和调节

磷酸化酶 (PE) 负责激活核苷/核苷酸逆转录酶抑制剂 (NRTI)，例如替诺福韦 (TFV)，并且对于它们转化以获得细胞内抗病毒活性至关重要。然而，关于 PE 的表达及其在女性生殖道中的活性的可用数据有限。这项工作比较了人类免疫缺陷病毒 (HIV) 研究中常用的人类女性生殖器组织、免疫细胞和动物模型中 PE 的信使 RNA (mRNA) 表达水平。此外，还评估了避孕激素和促炎细胞因子对人阴道、上皮和免疫细胞中替诺福韦二磷酸 (TFV-DP) 形成和功效的影响。我们发现人类阴道和外宫颈组织对于测试的七种 PE 具有相似的 mRNA 表达。聚合酶链反应结果显示，肌酸激酶脑 (CKB)、线粒体肌酸激酶 1 (CKMT1)、线粒体肌酸激酶 2 (CKMT2)、腺苷酸激酶 AK3L1 (AK4) 和核苷二磷酸激酶 1 (NME1) 表现出 10 至 10,000 -在阴道上皮细胞系 VK2 中的表达水平是 CD4 的两倍⁺ T 细胞 ( p  < .05)。醋酸甲羟孕酮 (MPA)/孕酮 (P4) 和 IL-1β/IL-8 处理导致 VK2 和 PM1 细胞中 TFV-DP 水平改变。当添加 1 μM TFV 时，浓度高于 0.1 μM 的 MPA 和 P4，以及浓度高于 10 ng/mL 的 IL-1β 和 IL-8，显着降低 PM1 细胞中的 HIV-1 _BaL抑制。然而，当 TFV 浓度升高到 1 mM 时，这种观察到的激素和细胞因子的作用被消除。这些体外结果阐明了 PE 在 TFV 代谢中的作用，并提供了有关 HIV 检测中常用动物模型的 PE 组织表达差异的信息。此信息可用于更好地理解和解释使用这些模型获得的数据。