Induction of Transient Virus Replication Facilitates Antigen-Independent Isolation of SIV-Specific Monoclonal Antibodies.
诱导瞬时病毒复制有利于 SIV 特异性单克隆抗体的抗原非依赖性隔离。
- 作者列表："Pedreño-Lopez N","Dang CM","Rosen BC","Ricciardi MJ","Bailey VK","Gutman MJ","Gonzalez-Nieto L","Pauthner MG","Le K","Song G","Andrabi R","Weisgrau KL","Pomplun N","Martinez-Navio JM","Fuchs SP","Wrammert J","Rakasz EG","Lifson JD","Martins MA","Burton DR","Watkins DI","Magnani DM
:Structural characterization of the HIV-1 Envelope (Env) glycoprotein has facilitated the development of Env probes to isolate HIV-specific monoclonal antibodies (mAbs). However, preclinical studies have largely evaluated these virus-specific mAbs against chimeric viruses, which do not naturally infect non-human primates, in contrast to the unconstrained simian immunodeficiency virus (SIV)mac239 clone. Given the paucity of native-like reagents for the isolation of SIV-specific B cells, we examined a method to isolate SIVmac239-specific mAbs without using Env probes. We first activated virus-specific B cells by inducing viral replication after the infusion of a CD8β-depleting mAb or withdrawal of antiretroviral therapy in SIVmac239-infected rhesus macaques. Following the rise in viremia, we observed 2- to 4-fold increases in the number of SIVmac239 Env-reactive plasmablasts in circulation. We then sorted these activated B cells and obtained 206 paired Ab sequences. After expressing 122 mAbs, we identified 14 Env-specific mAbs. While these Env-specific mAbs bound to both the SIVmac239 SOSIP.664 trimer and to infected primary rhesus CD4+ T cells, five also neutralized SIVmac316. Unfortunately, none of these mAbs neutralized SIVmac239. Our data show that this method can be used to isolate virus-specific mAbs without antigenic probes by inducing bursts of contemporary replicating viruses in vivo.
HIV-1 包膜 (Env) 糖蛋白的结构表征促进了分离 HIV 特异性单克隆抗体 (mab) 的 Env 探针的开发。然而，与无约束的猿猴免疫缺陷病毒 (SIV) 相比，临床前研究在很大程度上评估了这些针对嵌合病毒 es 的病毒特异性 mab，它们不会自然感染非人类灵长类动物 mac239 克隆。鉴于缺乏分离 SIV 特异性 B 细胞的天然样试剂，我们研究了一种无需使用 Env 探针分离 SIVmac239-specific mab 的方法。我们首先通过在 SIVmac239-infected 恒河猴输注 cd8 β 耗竭单克隆抗体或退出抗逆转录病毒治疗后诱导病毒复制来激活病毒特异性 b细胞。随着病毒血症的增加，我们观察到循环中 SIVmac239 Env 反应性浆细胞数量增加 2-4 倍。然后我们分选这些激活的 B 细胞，获得 206 个配对的 Ab 序列。表达 122 个 mab 后，我们鉴定出 14 个 Env 特异性 mab。虽然这些 Env 特异性 mab 与 SIVmac239 SOSIP.664 三聚体和感染的原代恒河猴 CD4 + T 细胞结合，但 5 个也中和了 sivmac316。不幸的是，这些 mab 都没有中和 sivmac239。我们的数据表明，这种方法可以通过诱导体内当代复制病毒 es 的爆发，在没有抗原探针的情况下分离病毒特异性 mab。
METHODS:BACKGROUND:Ultrasound has been demonstrated to accurately diagnose rectal deep endometriosis (DE) and pouch of Douglas (POD) obliteration. The role of ultrasound in the assessment of patients who have undergone surgery for rectal DE and POD obliteration has not been evaluated. AIM:To describe the transvaginal ultrasound (TVS) findings of patients who have undergone rectal surgery for DE. MATERIALS AND METHODS:An observational cross-sectional study at a tertiary care centre in Sydney, Australia between January and April 2017. Patients previously treated for rectal DE (low anterior resection vs rectal shaving/disc excision) were recruited and asked to complete a questionnaire on their current symptoms. On TVS, POD state and rectal DE were assessed. Correlating recurrence of POD obliteration and/or rectal DE to surgery type and symptoms was done. RESULTS:Fifty-six patients were contacted; 22/56 (39.3%) attended for the study visit. Average interval of surgery to study visit was 52.8 ± 24.6 months. Surgery type breakdown was as follows: low anterior resection (56%) and rectal shaving/disc excision (44%). The prevalence of POD obliteration was 16/22 (72.7%) intraoperatively and 8/22 (36.4%) at study visit, as per the sliding sign. Nine patients (39.1%) had evidence on TVS of recurrent rectal DE. Recurrence of POD obliteration and rectal DE was not associated with surgery type or symptomatology. CONCLUSION:Despite surgery for rectal DE, many patients have a negative sliding sign on TVS, representing POD obliteration, and rectal DE. Our numbers are too small to correlate with the surgery type or their current symptoms.
METHODS::Minimally invasive surgery for complex endometriosis requires preoperative planning that intimately connects the gynecologic surgeon to the radiologist. Understanding the surgeon's perspective to endometriosis treatment facilitates a productive relationship that ultimately benefits the patient. We examine minimally invasive surgery for endometriosis and the key radiologic information which enable the surgeon to successfully negotiate patient counseling, preoperative planning, and an interdisciplinary approach to surgery.
METHODS:STUDY OBJECTIVE:Prior research collectively shows that endometriosis is inversely related to women's adiposity. The aim of this study was to assess whether this inverse relationship holds true by disease severity and typology. DESIGN:Cross sectional study among women with no prior diagnosis of endometriosis. SETTING:Fourteen clinical centers in Salt Lake City, Utah and San Francisco, California. PATIENTS:Four hundred and ninety five women, ages 18-44 years, were enrolled in the operative cohort of the Endometriosis, Natural History, Diagnosis, and Outcomes (ENDO) Study. INTERVENTIONS:Gynecologic laparoscopy/laparotomy, regardless of clinical indication. MEASUREMENTS AND MAIN RESULTS:Participants underwent anthropometric assessments, body composition, and body fat distribution ratios before surgery. Surgeons completed a standardized operative report immediately after surgery to capture revised ASRM staging (I to IV) and typology of disease (superficial [SE], ovarian endometrioma [OE], and deep infiltrating endometriosis [DIE]). Linear mixed models, taking into account within-clinical-center correlation were used to generate least square means (95% confidence intervals) to assess differences in adiposity measures by endometriosis stage (no endometriosis, I-IV) and typology (no endometriosis, SE, DIE, OE, OE + DIE) adjusting for age, race/ethnicity, and parity. While the majority of confidence intervals were wide and overlapping, three general impressions emerged: 1) women with versus without incident endometriosis had the lowest anthropometric/body composition indicators; 2) women with stage I or IV had lower indicators compared to women with stage II or III; and 3) women with OE and/or DIE tended to have the lowest indicators, while women with SE had the highest indicators. CONCLUSION:Our research highlights that the relationship between women's adiposity and endometriosis severity and typology may be more complicated than prior research indicates.