Long-term vaginal antibody delivery: delivery systems and biodistribution

Topical delivery systems can provide prolonged delivery of antibodies to the vaginal mucosal surface for long-term protection against infectious diseases. We examined the biodistribution of antibodies during 30 days of vaginal antibody delivery in mice. Different antibody preparations (including monoclonal IgG and IgM, as well as several different (125)I-labeled IgGs) were administered by polymer vaginal rings, which were designed to provide continuous antibody delivery. Antibody concentrations remained high in the vaginal secretions for up to 30 days after disk insertion; radiolabeled antibody was also found, at approximately 100 times lower concentration, in the blood and other tissues. The measured concentrations agreed reasonably well with a simple pharmacokinetic model, which was used to calculate mucosal and systemic concentrations as a function of antibody delivery and elimination rates. Results from the model were consistent with previously reported antibody pharmacokinetic measurements: the half-life for antibody elimination for the vagina was approximately 3 h; the half-life for IgG(1) clearance from the blood was >1 day; and the overall permeability constant for vaginal uptake of IgG was approximately 0.01 to 0.03 h(-1). These results provide important information for the design of controlled antibody delivery devices for vaginal use, and suggest that high-dose, long-term vaginal administration of antibodies may be a reasonable approach for achieving sustained mucosal and systemic antibody levels.     

A Temperature-Monitoring Vaginal Ring for Measuring Adherence

BackgroundProduct adherence is a pivotal issue in the development of effective vaginal microbicides to reduce sexual transmission of HIV. To date, the six Phase III studies of vaginal gel products have relied primarily on self-reporting of adherence. Accurate and reliable methods for monitoring user adherence to microbicide-releasing vaginal rings have yet to be established.MethodsA silicone elastomer vaginal ring prototype containing an embedded, miniature temperature logger has been developed and tested in vitro and in cynomolgus macaques for its potential to continuously monitor environmental temperature and accurately determine episodes of ring insertion and removal.ResultsIn vitro studies demonstrated that DST nano-T temperature loggers encapsulated in medical grade silicone elastomer were able to accurately and continuously measure environmental temperature. The devices responded quickly to temperature changes despite being embedded in different thickness of silicone elastomer. Prototype vaginal rings measured higher temperatures compared with a subcutaneously implanted device, showed high sensitivity to diurnal fluctuations in vaginal temperature, and accurately detected periods of ring removal when tested in macaques.ConclusionsVaginal rings containing embedded temperature loggers may be useful in the assessment of product adherence in late-stage clinical trials.     

HIV-1 Specific IgA Detected in Vaginal Secretions of HIV Uninfected Women Participating in a Microbicide Trial in Southern Africa Are Primarily Directed Toward gp120 and gp140 Specificities

Background

Many participants in microbicide trials remain uninfected despite ongoing exposure to HIV-1. Determining the emergence and nature of mucosal HIV-specific immune responses in such women is important, since these responses may contribute to protection and could provide insight for the rational design of HIV-1 vaccines.

Methods and Findings

We first conducted a pilot study to compare three sampling devices (Dacron swabs, flocked nylon swabs and Merocel sponges) for detection of HIV-1-specific IgG and IgA antibodies in vaginal secretions. IgG antibodies from HIV-1-positive women reacted broadly across the full panel of eight HIV-1 envelope (Env) antigens tested, whereas IgA antibodies only reacted to the gp41 subunit. No Env-reactive antibodies were detected in the HIV-negative women. The three sampling devices yielded equal HIV-1-specific antibody titers, as well as total IgG and IgA concentrations. We then tested vaginal Dacron swabs archived from 57 HIV seronegative women who participated in a microbicide efficacy trial in Southern Africa (HPTN 035). We detected vaginal IgA antibodies directed at HIV-1 Env gp120/gp140 in six of these women, and at gp41 in another three women, but did not detect Env-specific IgG antibodies in any women.

Conclusion

Vaginal secretions of HIV-1 infected women contained IgG reactivity to a broad range of Env antigens and IgA reactivity to gp41. In contrast, Env-binding antibodies in the vaginal secretions of HIV-1 uninfected women participating in the microbicide trial were restricted to the IgA subtype and were mostly directed at HIV-1 gp120/gp140.     

Controlled vaginal delivery of antibodies in the mouse.

Controlled delivery of monoclonal antibodies to the mucus secretions of the vagina might provide women with passive immunoprotection against both sexually transmitted diseases and unwanted pregnancy. We have developed intravaginal devices composed of poly(ethylene-co-vinyl acetate) (EVAc) that continuously release IgG antibodies for over 30 days into buffered saline, and we have tested these devices in the vagina of mice. Polymeric devices containing either BSA (as a test reagent for proteins) or anti-hCG antibody, when inserted into the vaginas of mice, provided a continuous supply of either BSA or hCG-binding antibodies to the vaginal mucus for 30 days. Antibodies released by the devices achieved high concentration in the mucus within the lumen of the vagina, but did not significantly ascend into the uterine horns, as determined by epifluorescence microscopy of fluorescently labeled mouse IgG and by immunohistochemical localization of rabbit IgG. Our results suggest that long-term intravaginal delivery of functionally intact antibodies can be achieved with devices composed of EVAc.     

Development and evaluation of a vaginal ring device for sustained delivery of HIV microbicides to non-human primates

Background  There is considerable interest in developing coitally independent, sustained release formulations for long-term administration of HIV microbicides. Vaginal ring devices are at the forefront of this formulation strategy.
Methods  Non-medicated silicone elastomer vaginal rings were prepared having a range of appropriate dimensions for testing vaginal fit in pig-tailed and Chinese rhesus macaques. Cervicovaginal proinflammatory markers were evaluated. Compression testing was performed to compare the relative flexibility of various macaque and commercial human rings.
Results  All rings remained in place during the study period and no tissue irritation or significant induction of cervicovaginal proinflammatory markers or signs of physical discomfort were observed during the 8-week study period.
Conclusions  Qualitative evaluation suggests that the 25 × 5-mm ring provided optimal fit in both macaque species. Based on the results presented here, low-consistency silicone elastomers do not cause irritation in macaques and are proposed as suitable materials for the manufacture of microbicide-loaded vaginal rings.     

Distribution of Candida species in different clinical sources in Delhi, India, and proteinase and phospholipase activity of Candida albicans isolates

Eighty-five isolates of Candida recovered from three hundred and fifty diverse clinical sources, viz. respiratory tract (sputum, bronchial washing,bronchoalveolar lavage, tracheal aspirate), blood, urine, high vaginal swab, skin and plastic devices, were studied in detail for their morphological and biochemical characters. Seven species of Candida were identified, viz., C. albicans (45.8%), C. tropicalis (24.7%), C. parapsilosis (10.5%), C. krusei (7.0%), C. kefyr (7.0%), C. guilliermondii (3.5%), and C. glabrata (1.1%). C. albicans was the predominant species isolated from all clinical specimens, except blood from which C. krusei was most frequently (38.4%) recovered. Out of 39 isolates of C. albicans, 26 (66.6%) and 19 (48.7%) exhibited strong proteinase and phospholipase activity respectively. There was a higher prevalence of proteinase producing strains amongst the vaginal and skin isolates than that in urinary and respiratory isolates. Also a greater number of phospholipase producing strains was observed in the vaginal and urinary isolates than that in the respiratory and skin isolates.     

Investigation of the Physicochemical and Physicomechanical Properties of a Novel Intravaginal Bioadhesive Polymeric Device in the Pig Model

The purpose of this study was to develop and evaluate the bioadhesivity, in vitro drug release, and permeation of an intravaginal bioadhesive polymeric device (IBPD) loaded with 3′-azido-3′-deoxythymidine (AZT) and polystyrene sulfonate (PSS). Modified polyamide 6,10, poly(lactic-coglycolic acid), polyacrylic acid, polyvinyl alcohol, and ethylcellulose were blended with model drugs AZT and PSS as well as radio-opaque barium sulfate (BaSO4) and then compressed into caplet devices on a tableting press. One set of devices was coated with 2% w/v pentaerythritol polyacrylic acid (APE-PAA) while another remained uncoated. Thermal analysis was performed on the constituent polymers as well the IBPD. The changes in micro-environmental pH within the simulated human vaginal fluid due to the presence of the IBPD were assessed over a period of 30 days. Textural profile analysis indicated that the bioadhesivity of the APE-PAA-coated devices (3.699 ± 0.464 N; 0.0098 ± 0.0004 J) was higher than that of the uncoated devices (1.198 ± 0.150 N; 0.0019 ± 0.0001 J). In addition, BaSO4-facilitated X-ray imaging revealed that the IBPD adhered to pig vaginal tissue over the experimental period of 30 days. Controlled drug release kinetics was obtained over 72 days. During a 24-h permeation study, an increase in drug flux for both AZT (0.84 mg cm⁻² h⁻¹) and PSS (0.72 mg cm⁻² h⁻¹) was realized up to 12 h and thereafter a steady-state was achieved. The diffusion and dissolution dynamics were mechanistically deduced based on a chemometric and molecular structure modeling approach. Overall, results suggested that the IBPD may be sufficiently bioadhesive with desirable physicochemical and physicomechanical stability for use as a prolonged intravaginal drug delivery device.     

1500例女性阴道微生态状况分析

目的女性阴道微生态状况调查与分析。方法采用阴道pH、阴道涂片Gram染色镜检(菌群密集度、菌群多样性、优势菌、真菌孢子、滴虫和炎性反应等)、阴道微生物功能测定,评估1 500例体检者阴道微生态状况。结果微生态正常830例(55.7%),微生态失调者670例(44.6%)。在670例微生态失调患者中,需氧菌阴道炎156例(23.3%),细菌性阴道病265例(39.6%),外阴阴道假丝酵母菌病31例(4.63%),滴虫性阴道炎2例(0.29%),混合感染216例(32.2%)。结论阴道微生态评价有助于筛查无症状阴道微生态失调患者,对指导临床评价感染及预防阴道感染具有重要的临床意义。     

Fertility, ovulation and maturation of eggs in mares injected with HCG

Pony mares were observed from January to August for incidence of oestrus, duration of oestrus, length of the oestrous cycle and for ovulation and fertility after injection of HCG. From January to 15 May most mares showed oestrus but the duration of oestrus was quite variable and few mares ovulated in response to HCG. From 15 May to 17 August oestrous cycles were more regular and ovulation was induced within 40-50 h by an intramuscular injection of 1500-5000 i.u. HCG. Pregnancy was established by one mating at a fixed time after HCG in 20 of 69 mares. Degenerate eggs were recovered from the oviducts of anoestrous recently ovulated, mated, unmated and pregnant mares. The first polar body was formed before ovulation in 2 eggs and had not formed in 2 recently ovulated eggs flushed from the oviduct. The second polar body formed after sperm penetration 10-12 h after ovulation. After formation of pronuclei, the first cleavage division occurred at 20 h and the second at 32 h after ovulation. Oestrus was inhibited by progesterone administered by vaginal devices but occurred within 1-3 days in 12 of the 20 mares after withdrawal of the devices.     

Genotype effects on body temperature in dairy cows under grazing conditions in a hot climate including evidence for heterosis

We compared diurnal patterns of vaginal temperature in lactating cows under grazing conditions to evaluate genotype effects on body temperature regulation. Genotypes evaluated were Holstein, Jersey, Jersey × Holstein and Swedish Red × Holstein. The comparison of Holstein and Jersey versus Jersey × Holstein provided a test of whether heterosis effects body temperature regulation. Cows were fitted with intravaginal temperature recording devices that measured vaginal temperature every 15 min for 7 days. Vaginal temperature was affected by time of day (P < 0.0001) and genotype × time (P < 0.0001) regardless of whether days in milk and milk yield were used as covariates. Additional analyses indicated that the Swedish Red × Holstein had a different pattern of vaginal temperatures than the other three genotypes (Swedish Red × Holstein vs others × time; P < 0.0001) and that Holstein and Jersey had a different pattern than Jersey × Holstein [(Holstein + Jersey vs Jersey × Holstein) × time, P < 0.0001]. However, Holstein had a similar pattern to Jersey [(Holstein vs Jersey) × time, P > 0.10]. These genotype × time interactions reflect two effects. First, Swedish Red × Holstein had higher vaginal temperatures than the other genotypes in the late morning and afternoon but not after the evening milking. Secondly, Jersey × Holstein had lower vaginal temperatures than other genotypes in the late morning and afternoon and again in the late night and early morning. Results point out that there are effects of specific genotypes and evidence for heterosis on regulation of body temperature of lactating cows maintained under grazing conditions and suggest that genetic improvement for thermotolerance through breed choice or genetic selection is possible.     

600例健康体检妇女阴道微生态状况分析

目的研究深圳宝安地区健康妇女阴道微生态状况。方法对2011年9月至2012年2月在深圳宝安区人民医院体检中心进行健康体检的600例妇女阴道分泌物进行检测,检测指标包括清洁度、念珠菌、滴虫、阴道酸碱度、唾液酸苷酶、白细胞酯酶、过氧化氢（H2O2）浓度。结果研究对象中阴道微生态正常328例（54．67％）,微生态失调者272例（45．33％）;阴道微生态失调者中,微生态功能检测阳性率及致病菌的检出率均明显大于微生态正常者,差异有统计学意义（P〈0．05）。结论深圳宝安地区健康体检妇女部分患有阴道炎或存在阴道微生态失调或亚失调状态;阴道微生态联合检测法简便、快速、价廉实用,值得推广。     

482例产褥期阴道微生态调查

目的了解产褥期妇女阴道微生态状况,为临床保健提供循证依据。方法选择长兴县妇幼保健院产后门诊复查的产褥期妇女482例及来院体检的健康妇女86例,采集阴道分泌物作pH、病原体、清洁度和阴道微生态功能检查,进行阴道微生态学评价、比较。结果 482例产褥期阴道pH为(4.62±0.62);阴道清洁度Ⅲ~Ⅳ127例;微生态正常者75例,占15.56%;微生态失调者407例,占84.43%。86例健康妇女阴道pH(4.38±0.65);阴道清洁度Ⅲ~Ⅳ14例;微生态正常者34例,占39.53%;微生态失调者52例,占60.47%。差异有统计学意义。结论产褥期妇女阴道微生态发生改变,表现为pH升高,清洁度下降,微生态失调比例明显升高,应引起临床保健工作者关注。     

True vaginal prolapse in a bitch

Frequently, vaginal fold prolapse is the protrusion of edematous vaginal tissue into and through the opening of the vulva occurring during proestrus and estrus stages of the sexual cycle. True vaginal prolapse may occur near parturition, as the concentration of serum progesterone declines and the concentration of serum oestrogen increases. In the bitch, this type of true vaginal prolapse is a very rare condition.

This short communication describes a 5-year-old female, cross-breed dog in moderate condition, weighing 33 kg, with distocia and true vaginal prolapse. Abdominal palpation and transabdominal ultrasonography revealed live and dead foetuses in the uterine horns. One dead and four live fetuses were removed from uterus by cesarean section. The ovariohysterectomy was performed after repositioning the vaginal wall with a combination of traction from within the abdomen and external manipulation through the vulva. Re-occurrence of a vaginal prolapse was not observed and the bitch recovered completely after the surgical therapy.

Compared to other vaginal disorders, vaginal prolapse is an uncommon condition in the bitch. In the present case, extreme tenesmus arising from distocia may have predisposed to the vaginal prolapse. The cause of dystocia was probably the disposition of the first foetus. We concluded that the vaginal prolapse was the result of dystocia in the present case.     

Effects of pelvic floor muscle contraction on anal canal pressure

The role of pelvic floor muscle contraction in the genesis of anal canal pressure is not clear. Recent studies have suggested that vaginal distension increases pelvic floor muscle contraction. We studied the effects of vaginal distension on anal canal pressure in 15 nullipara asymptomatic women. Anal pressure, rest, and squeeze were measured using station pull-through manometry techniques with no vaginal probe, a 10-mm vaginal probe, and a 25-mm vaginal probe in place. Rest and squeeze vaginal pressures were significantly higher when measured with the 25-mm probe compared with the 10-mm probe, suggesting that vaginal distension enhances pelvic floor contraction. In the presence of the 25-mm vaginal probe, rest and squeeze anal pressures in the proximal part of the anal canal were significantly higher compared with no vaginal probe or the 10-mm vaginal probe. On the other hand, distal anal pressures were not affected by any of the vaginal probes. Ultrasound imaging of the pelvic floor revealed that vaginal distension increased the anterior-posterior length of the puborectalis muscle. Atropine at 15 micro g/kg had no influence on the rest and squeeze anal pressures with or without vaginal distension. Our data suggest that pelvic floor contractions increase pressures in the proximal part of the anal canal, which is anatomically surrounded by the puborectalis muscle. We propose that pelvic floor contraction plays an important role in the fecal continence mechanism by increasing anal canal pressure.     

Adhesiveness of yeast-like fungi in the system "Candida albicans--vaginal epitheliocytes"

The influence of vaginal liquid on adhesive reactions in the system "Candida albicans - vaginal epitheliocytes". Preliminary treatment of C. albicans by mucin, natural or absorbed vaginal liquid resulted in decrease of adhesive activity (p<0.05). C. albicans did not change its adhesive properties in case of preliminary treatment of vaginal epitheliocytes by the same substances. In the system "Candida albicans - vaginal epitheliocytes" vaginal liquid has the expressed antiadhesive effect, which probably belongs to mucin.     

不同生理阶段、避孕方式和感染状态对阴道乳酸杆菌影响的研究

目的调查不同生理阶段、避孕方式、感染因素对女性阴道乳酸杆菌数量的影响。方法收集南昌大学第二附院妇产科就诊的176例妇女的主要资料并采用qPCR方法定量分析阴道分泌物中乳酸杆菌数量,分泌物涂片和革兰染色后在显微镜下观察记录其感染状态。结果非绝经期组不同年龄段乳酸杆菌数量差异无统计学意义,绝经期组乳酸杆菌数量减少;不同避孕方式组的阴道乳酸杆菌数量差异无统计学意义;细菌性阴道病和假丝酵母菌感染组阴道乳酸杆菌数量较正常组差异均有统计学意义。结论生理阶段、细菌性阴道病和假丝酵母菌感染是影响女性阴道乳酸杆菌数量的主要因素。     

Age- and hormone-related changes in vaginal smear patterns in the gray-tailed vole, Microtus canicaudus

Female voles, Microtus canicaudus, exhibited age-related changes in vaginal smear patterns when isolated from males after weaning. Between 30 and 50 days of age, nearly all females exhibited persistently leucocytic vaginal smears. By 90-120 days, most females showed vaginal cyclicity with alternating predominance of leucocytes, nucleated epithelial cells or cornified epithelial cells. Most females examined between 150 and 200 days of age exhibited persistent vaginal cornification. The vaginal cyclicity seen in females between 90 and 120 days was not a reflection of cyclic ovulatory changes; plasma progesterone concentrations remained constant, regardless of age or vaginal smear pattern, and corpora lutea were never seen in unmated females. Although progesterone concentrations did not differ among vaginal smear patterns of 120-day-old females, plasma oestrogen values were highest in females exhibiting vaginal cornification.     

4019例妇科门诊不同症状患者阴道微生态状况分析

目的探讨妇科门诊不同症状患者的阴道微生态状况。方法回顾性分析2011年11月至2012年9月西安交通大学医学院第一附属医院妇科门诊因不同症状就诊患者的阴道微生态评价（菌群的密集度、多样性、优势菌、病原菌、AV评分、Nugent评分及五项菌群功能及炎症反应指标）检测结果。结果4019例被检查者平均年龄为33．56岁。其中阴道微生态正常占n70％（28／4019）;阴道微生态失调者占99．30％（3991／4019）;后者包括未明确感染的阴道微生态失调患者占66．36％（2667／4019）;明确感染的阴道微生态失调患者占32．94％（1324／4019）;其中外阴阴道假丝酵母菌病（vulvovaginal candidiasis,VVC）患者占24．33％（978／4019）;需氧菌陛阴道炎（aerobic vaginitis,AV）患者占3．11％（125／4019）;细菌性阴道病（bacterial vaginosis,BV）患者占3．14％（126／4019）;滴虫阴道炎（trichomonas vaginitis,TV）患者占5．15％（207／4019）;混合感染患者占8．31％（110／1324）。优势菌群异常患者占54．32％（2183／4019）。结论妇科门诊中未明确感染的阴道微生态失调患者构成比最高;在明确感染的患者中外阴阴道假丝酵母菌病构成比最高;因此阴道微生态评价成为阴道感染性疾病诊断首选检测方法,在临床诊疗实践中具有重要意义。     

Development of a self-contained,indwelling vaginal temperature probe for use in cattle research

A device was developed to monitor the vaginal temperature of cattle in a research setting. This device decreases labor involved with monitoring body temperature compared with manual temperature readings, allows for continuous monitoring of vaginal temperature at 1 min intervals, and also allows for temperature measurements without the presence of a human handler or without restraint, which can agitate cattle. The device consists of a blank controlled internal drug release (CIDR) device (designed by Pfizer Animal Health as an indwelling vaginal probe) that holds an indwelling vaginal temperature probe logger. The fabrication of the vaginal probe costs approximately US $325 per unit. Similar rectal and vaginal temperature responses to lipopolysaccharide challenge were observed when vaginal and rectal temperatures were measured simultaneously in the same heifer (P>0.05). Additionally, rectal and vaginal temperatures were highly correlated (r=0.97; P<0.0001). Similar to the rectal temperature monitoring device, the vaginal device allows for the measurement of vaginal temperature without the potential biases associated with the stress response produced as a reaction to the handling by and (or) presence of humans. The vaginal temperature recording device will provide researchers with an additional inexpensive tool to study physiological responses in female cattle.     

纵向研究妊娠期妇女的阴道微生态状况

目的分析妊娠期妇女阴道微生态状况的纵向变化。方法选择健康无症状单胎妊娠期妇女50例,分别于孕早期（10.2±2.1）周、孕中期（24.6±3.7）周、孕晚期（33.2±3.4）周,取其阴道分泌物,进行pH、H2O2含量检测及细菌培养,并经革兰染色后在油镜下观察,进行阴道微生态（阴道菌群的密集度、多样性、优势菌、炎症反应状况等）状况评价。结果 （1）50例孕妇中,孕早期38例（76%）阴道微生态正常,其中31例（81.58%）整个孕期维持阴道微生态正常,7例（18.42%）孕妇于孕中期或孕晚期转变为阴道微生态失调;12例（24%）孕早期阴道微生态失调,其中6例（50%）于孕中期或孕晚期转变为阴道微生态正常。（2）无症状性细菌性阴道病随着孕周增加部分可自然消退40%（2/5）。（3）H2O2阳性、阴道pH低者易从阴道微生态失调转为阴道微生态正常。结论妊娠期阴道微生态是动态变化的,部分细菌性阴道炎可自然消退,H2O2阳性及低阴道pH为维持正常阴道微生态的重要因素。