洛基维特单抗治疗对特应性皮炎犬经表皮失水量的影响

王帆

The influence of treatment with lokivetmab on transepidermal water loss (TEWL) in dogs with spontaneously occurring atopic dermatitis

洛基维特单抗治疗对特应性皮炎犬经表皮失水量的影响

翻译：杨益蓓 校对：王帆

Background – Atopic dermatitis (AD) is one of the most common skin diseases in dogs. Monitoring the progress of treatment may include assessment of lesion severity by use of Canine Atopic Dermatitis Extent and Severity Index (CADESI), assessment of pruritus and measurement of biophysical parameters of the skin.

Hypothesis – This study aimed to assess changes in transepidermal water loss (TEWL) during 12 weeks of treatment with lokivetmab and its correlation with the CADESI-04 and the severity of pruritus.

Animals – Ten client-owned dogs with canine AD.

Methods and materials – The animals were administered lokivetmab three times at four week intervals. Pruritus and CADESI-04 were assessed, and TEWL was measured at six body regions, on Day 0 and repeated at weeks 4, 8 and 12.

Results – There was a significant decrease in the CADESI-04 score, the severity of pruritus and the mean TEWL between baseline and each of the time points. TEWL also decreased in the majority of body sites examined. Significant correlations between the CADESI-04 and mean TEWL (r = 0.519, P = 0.0003), and between pruritus severity and mean TEWL (r = 0.517, P = 0.0003), were found.

Conclusions and clinical significance – Overall TEWL decreased during lokivetmab treatment but not at all body sites. If TEWL is used to assess the effectiveness of a treatment, a mean TEWL score should be calculated from several areas of the body.

摘要

背景-特应性皮炎(AD)是犬最常见的皮肤病之一。监测治疗进展包括评估犬特应性皮炎严重程度指数(CADESI)、 瘙痒评分和皮肤生物物理参数,进而评估病变的严重程度。

假设-本研究洛基维特单抗12周的治疗,旨在评估在此期间犬经表皮失水量(TEWL)的改变,及其与cadesi-04和瘙痒程度的相关性。

动物-十只患有特应性皮炎的家养犬。

方法和材料-每隔四周给动物注射1次洛基维特单抗,连续3次。对瘙痒和CADESI-04进行评分,并分别在第0天、第4周、第8 周和第12周测量6个机体部位的TEWL。

结果-与基础值相比,其他每个时间点的CADESI-04分值、瘙痒程度和平均TEWL都显著降低了。大多数被检测机体部位的TEWL也下降了。在CADESI-04和平均TEWL(r=0.519,p=0.0003)之间、以及瘙痒程度和平均TEWL(r=0.517,p=0.0003)之间存在显著相关性。

结论和临床意义-在洛基维特单抗治疗期间,整体TEWL降低,但并非所有机体部位都降低了。如果使用TEWL来评估治疗的有效 性,则应计算若干机体部位的平均TEWL。

Introduction

介绍

Atopic dermatitis (AD) is a common allergic skin disease in dogs and the pathogenesis has been associated with disturbances in skin barrier function.The assessment of skin barrier function, using biophysical methods, may include measurement of epidermal hydration, erythema, skin pH and, most commonly, transepidermal water loss (TEWL). Increased transepidermal water loss was reported in the course of allergic disease in dogs. Treatments, such as ciclosporin and allergen-specific immunotherapy, have been associated with a reduction in values for TEWL. During treatment, a decrease in TEWL value was observed for various areas of the body. Some studies have reported correlation between TEWL values, for some of the assessed body areas, and clinical assessment systems including the Canine Atopic Dermatitis Extent and Severity Index – CADESI.

特应性皮炎(AD)是犬常见的过敏性皮肤病，其发病机制与皮肤屏障功能紊乱有关。皮肤屏障功能的评估，使用生物物理方法，可包括测量表皮水合、皮肤发红、皮肤pH和最常见的经表皮失水量(TEWL)。犬特应性皮炎的病程中有报道经表皮失水量增加。治疗，如环孢素和过敏原特异性免疫治疗，与经表皮失水量值的降低有关。治疗期间，在机体各个区域可观察到经表皮失水量值的降低。一些研究报告了若干被评估的机体区域的经表皮失水量值与包括犬特应性皮炎的严重程度指数-CADESI在内的临床评估系统之间的相关性。

Lokivetmab (Cytopoint, Zoetis Belgium SA; Louvain-laNeuve, Belgium) is a caninized anti-IL-31 antibody that binds circulating IL-31 and prevents its binding to the receptor. The subcutaneous (s.c.) administration of lokivetmab to atopic dogs has been reported to reduce pruritus for four weeks and in the improvement of skin lesions, based on the assessment of the third iteration of CADESI(-03).

洛基维特单抗（赛妥敏）是一种犬化的抗IL-31抗体，与游离IL-31结合，阻止其与受体结合。基于第三代犬特应性皮炎严重程度指数的评估系统，皮下注射洛基维特单抗被报道可减少瘙痒持续四周和改善皮肤病变。

This aim of this study was to assess TEWL in atopic dogs over the course of 12 weeks of treatment with lokivetmab and its correlation with the CADESI-04 scoring system and an evaluation of the severity of pruritus.

本研究的目的是在洛基维特单抗治疗12周期间，评估特应性皮炎犬TEWL与CADESI-04评分系统和瘙痒程度的相关性。

Methods and materials

方法和材料

Animals

动物

Ten dogs were recruited (five male and five female), aged from nine months to six years (median 16 months) weighing from 8.5 to 40 kg (median 11.65 kg). Dog breeds included French bulldog (four individuals), akita inu, fox terrier, West Highland white terrier, Labrador retriever, Boston terrier and cross-breed. The dogs were privately owned and were presented to the Department of Clinical Diagnostics and Veterinary Dermatology at the University of Life Sciences in Lublin. Owners’ written consent to participate in the study was obtained before treatment. All procedures used in this study (TEWL measurement, clinical assessment of the CADESI and pruritus scores) were noninvasive. The Local Ethics Board in Lublin considered all procedures to be routine veterinary and did not require institutional approval. Lokivetmab was used according to manufacturer’s recommendation. The characteristics of the dogs included in the study are reported in Table 1.

招募了10只犬（5只雄性和5只雌性)，年龄从9个月到6岁(中位数16个月)，体重从8.5公斤到40公斤(中位数11.65公斤）。犬种包括法国斗牛犬（4只）、秋田犬、狐狸㹴、西高地白㹴、拉布拉多猎犬、波士顿猎犬和杂交品种。这些犬是家养犬，就诊于卢布林生命科学大学临床诊断和兽医皮肤科。在治疗前获得宠主书面同意参加这项研究。本研究中使用的所有程序(TEWL测量、CADESI临床评估和瘙痒评分)都是非侵入性的。卢布林的地方道德委员会认为所有程序都是例行的兽医程序，不需要机构批准。洛基维特单抗是根据制造商的建议使用的。研究中包括的犬的特征见表1。

表1研究开始时10只犬的特征和基线评估数据（第0天）

Dogs were diagnosed with AD on the basis of published diagnostic criteria (set 2). Other pruritic diseases were excluded based on trichoscopic examination, skin scrapings and cytological evaluation. Flea bite hypersensitivity and food allergy were excluded by appropriate anti-flea treatments and a strict eight week elimination diet. Dogs were sampled for a commercial test reporting allergen-specific IgE antibodies (PolyCheck Allergies Dog Panel, BioCheck GmBH; Munster, Germany). Sampling was performed during a period of clinical flare before treatment. The dogs were not treated before the study and did not receive ciclosporin, oclacitinib, glucocorticoids, antihistamines, polyunsaturated fatty acids, pentoxifylline and lokivetmab for at least two months before the study.

根据已公布的诊断标准（第2套），犬被诊断为特应性皮炎。其他瘙痒性疾病根据毛发检查，皮肤刮片和细胞学评估排除。跳蚤叮咬过敏和食物过敏通过适当的抗跳蚤治疗和严格的八周食物排查试验被排除。对犬进行抽样检测，报告过敏原特异性IgE抗体。在治疗前的临床爆发期间进行取样。在研究之前，犬没有接受治疗，在研究前至少2个月没有使用环孢素、奥拉替尼、糖皮质激素、抗组胺药、多不饱和脂肪酸、己酮可可碱和洛基维特单抗。

Lokivetmab treatment

洛基维特单抗治疗

The dogs were treated with lokivetmab (Cytopoint, Zoetis) using 1 mL single-use vials containing 10, 20, 30 or 40 mg/mL. Dogs weighing ≤10 kg received 10 mg lokivetmab, 10–20 kg dogs received 20 mg, 20–30 kg dogs received 30 mg and 30–40 kg dogs received 40 mg, according to the manufacturer’s instructions. The dose of the drug administered ranged from 1 to 1.76 mg/kg (mean 1.26 mg/kg) depending on the weight of the animal. Lokivetmab was administered s.c. three times at four week intervals.

用含有10、20、30或40mg/ml的1ml单用小瓶对犬进行洛基维特单抗(赛妥敏)治疗。根据制造商的指示，体重≤10kg的犬接受10mg洛基维特单抗，10-20kg的犬接受20mg，20-30kg的犬接受30mg，30-40kg的犬接受40mg。根据动物的体重，给药剂量1至1.76mg/kg(平均1.26 mg/kg）。洛基维特单抗皮下注射每隔四周给药一次，共给药三次。

TEWL measurement

经表皮失水量测量

Measurements of TEWL were performed in dogs before treatment (Week 0) and three times at four week intervals (at weeks 4, 8, and12). Measurements were made on six areas of the body including the concave surface of the right auricle, interdigital space of the right forelimb, right lateral thoracic area, abdomen, right axilla and right groin. The measurements were obtained after acclimatization of the animals in the room for ≥60 min. The temperature in the room where the test was carried out ranged from 21 to 25°C and humidity ranged from 41 to 60%. If necessary (for the lateral thoracic area), the hairs were clipped using an electric razor (Type 1245, 0.3 mm blade, Moser Animalline; Unterkirnach, Germany) before the measurement. The hairs in nonhairless areas of the body (auricle, abdomen, axilla, groin and interdigital space) were not clipped. Measurement of TEWL was performed using a vapometer with a closed chamber (VapoMeter SWL 4605, DELFIN Technologies; Kuopio, Finland). Each measurement was performed three times by the same operator and the average of three measurements was used for further analysis.

在治疗前（第0周)和每隔四周1次共3次(第4、8和12周）对犬进行TEWL测量。测量机体的六个区域，包括右耳的凹面、右前肢的指间隙、右侧胸区、腹部、右腋窝和右腹股沟。动物在室内适应至少60分钟后测量。进行测试的房间的温度在21到25°C之间，湿度在41%到60%之间。如有必要（对于侧胸区），在测量之前，用电动剃须刀剃毛。机体无毛区（耳廓、腹部、腋窝、腹股沟和指间间隙）的毛发未被修剪 TEWL的测量是在密闭室内使用蒸气计进行的 每个测量由同一操作者进行三次，并使用三次测量的平均值进行进一步分析。

CADESI-04 and numerical pruritus scale (NPS)

犬特应性皮炎严重程度指数第四版和瘙痒程度得分

For each dog at each visit, the CADESI-04 scoring system was applied immediately before TEWL determination.The severity of pruritus was assessed using a numerical six point scale from 0 to 5 based on a previously published method.

对每次查看的每一只犬，在TEWL测定前应立即应用CADESI-04评分系统。 根据先前发表的方法，使用从0到5的数值六点量表评估瘙痒的严重程度。

Statistical analysis

统计分析

Normal distribution of the obtained results was tested with the Shapiro–Wilk test. Statistical differences between individual measurements at weeks 0, 4, 8 and 12 for the CADESI and severity of pruritus were calculated. Each dog’s average TEWL (aTEWL) was calculated and counted as the mean of six measurements on each area of the body in weeks 0, 4, 8 and 12. The differences between measurements at weeks 0, 4, 8 and 12 for aTEWL were calculated. The differences in TEWL between measurements at weeks 0, 4, 8 and 12 also were calculated separately for each of the six assessed body regions. The differences in statistical significance between the results were calculated based on the Wicoxon–Mann–Whitney U-test. In all cases Bonferroni corrections were implemented. P < 0.05 was considered significant. Additionally, the correlation coefficients between aTEWL and CADESI-04, aTEWL and NPS, and CADESI-04 and TEWL on individually assessed regions, and between NPS and TEWL on particular assessed regions were calculated. Spearman test for P < 0.05 was used to calculate the correlation. All analyses were performed using Statistica 10 software (Statsoft; Tulsa, OK, USA).

所得结果的正态分布用夏皮罗-威尔克检验进行了检验。计算了0、4、8、12周个体测量的CADESI和瘙痒程度的统计学差异。在0、4、8和12周每个机体部位6次测量的平均值被作为每只犬的平均TEWL(aTEWL)被计算出来。计算了0、4、8、12周aTEWL的测量值差异。也分别计算了6个被评估机体区域中的每个区域在0、4、8和12周的TEWL测量值之间的差异。结果之间统计学意义上的差异是根据 wioxon-mann-whitney u-test 计算出来的。在所有情况下，Bonferroni校正被执行。P < 0.05被认为是显著的。此外计算了aTEWL与CADESI-04，aTEWL与NPS，CADESI-04与TEWL在各个评估区域的相关性，NPS与TEWL在特定的评估区域的相关性。 采用P<0.05的Spearman检验计算相关性。 所有分析均使用Statistica10软件进行。

图1犬特应性皮炎严重程度指数(CADESI)与平均经表皮失水量(aTEWL)的相关性。

实线，趋势线；虚线，0.95系数区间。

图2瘙痒量化表(NPS)与平均经皮失水(aTEWL)之间的相关性。

实线，趋势线；虚线，0.95系数区间。

Results

结果

Signalment and initial values for CADESI, NPS and TEWL (at Day/Week 0) for 10 dogs are presented in Table 1.

表1列出了10只犬的CADESI、NPS 和TEWL (在第0天/周)的特征和初始值。

During therapy, gradual and statistically significantly decreases in CADESI-04 scores, pruritus measurements and overall TEWL were observed after four weeks; reductions were not statistically significant after eight and 12 weeks of therapy (Table 2). Changes were also noted for TEWL at individual body sites, some of which were statistically significant (Table 3).

在治疗期间，4周之后观察到CADESI-04评分，瘙痒测量和整体TEWL逐渐下降，且有统计学意义； 治疗8周和12周后，减少量无统计学意义（表2）。个别机体部位的TEWL也有变化，其中一些有统计学意义（表3）。

表2在使用洛基维特单抗治疗12周的犬，犬特应性皮炎严重程度指数，第4代(CADESI-04)评分，瘙痒测量和平均经表皮失水量

表3接受洛基维特单抗治疗12周特应性皮炎犬6个体区的经表皮失水量(TEWL)值

A significant correlation was found between aTEWL and CADESI-04 (r = 0.519, P = 0.0003) and between aTEWL and NPS (r = 0.517, P = 0.0003). The results for the correlations are shown in Figures 1 and 2. Correlations between CADESI-04 and TEWL, and between NPS and TEWL, at individual body regions are reported in Table S1.

aTEWL与CADESI-04(r = 0.519, P = 0.0003)，aTEWL与NPS (r = 0.517, P = 0.0003)之间发现了显著的相关性。图表1,2给出了相关的结果。表S1报告了CADESI-04和TEWL之间的相关性，以及NPS和TEWL之间的相关性。

Discussion

讨论

Lokivetmab administered s.c. to a group of atopic dogs was associated with significant reductions in the clinically evaluated CADESI-04 parameters, pruritus severity and average TEWL values after four weeks. There also were gradual reductions in TEWL in individual areas of the body, although varying intensities were noted depending on the area assessed. For example, values for the abdomen were statistically different between the time points, similar to findings for the aTEWL. There were no significant differences for any of the measurements for the thoracic area. It is difficult to explain why a decrease in TEWL was not observed in all examined body areas. Reductions in this parameter during treatment were observed in body areas commonly associated with AD lesions, including the abdomen, interdigital area, axilla and groin, but not in the thorax, which is not a common site for lesions of AD. These results suggest that the former body areas should be exclusively examined in future research.

洛基维特单抗用于一组特应性皮炎犬四周后，临床评估的CADESI-04参数，瘙痒的严重程度和平均经表皮失水量显著降低。在机体的各个部位，TEWL也逐渐减少，虽然变化的强度取决于评估的区域。例如，不同时间点之间，腹部的数值有统计学上的差异，类似于aTEWL的结果。胸部区域的任何测量结果均无显着性差异。很难解释为什么没有在所有被检查的机体区域观察到TEWL的下降。在治疗过程中，这一参数的减少通常在与特应性皮炎病变有关的机体部位，包括腹部、趾间区、腋下和腹股沟，但在不是特应性皮炎病变常见部位的胸部没有减少。这些结果表明，以上的机体区域应该在未来的研究中专门检查。

There appeared to be a correlation between values for CADESI-04 and TEWL, which was most evident for the average TEWL. Correlations between CADESI-04 and TEWL for individual body areas were weaker and significant for only half of the body areas examined. Likewise, when assessing the correlation between aTEWL and the severity of pruritus, the correlation was stronger than those observed between the severity of pruritus and TEWL assessed at individual body areas. We recommend that aTEWL should be used in future studies to assess skin barrier function.

CADESI-04和TEWL的值之间似乎存在相关性，这在aTEWL中最为明显。CADESI-04和TEWL在单个机体区域的相关性较弱，仅在被检查的一半机体区域有显着性差异。同样地，当评估aTEWL和瘙痒严重程度之间的相关性时，这种相关性比在个别机体部位的瘙痒严重程度和TEWL之间的相关性强。 我们建议在未来的研究中使用aTEWL来评估皮肤屏障功能。

To the best of the authors’ knowledge, TEWL has not been used to assess the effectiveness of treatment with lokivetmab. A previous study reported a decrease in the CADESI-03 value and pruritus after the use of lokivetmab and this was most apparent with the single application of a dose of two mg/kg.The duration of the effect on pruritus was 49 days and for CADESI-03 it was eight weeks. Another study evaluated the effectiveness of lokivetmab administered monthly using CADESI-03 and the Pruritus Visual Analog Scale (PVAS); there was a significant reduction in the CADESI-03 (from 184 to 57) after 12 weeks of treatment.The decrease was greatest after the first administration of the drug (CADESI-03 reduced from 184 to 76 after four weeks; 54% reduction). This finding is similar to our study where the largest decrease, in CADESI-04, was also recorded after four weeks (from 36.6 to 11.1; 69% reduction). For pruritus, the greatest decrease was observed after four weeks, a reduction of greater than 51% compared with a 64% decrease in our study.

据作者所知，TEWL还没有被用来评估洛基维特单抗治疗的有效性。先前的一项研究报告，使用洛基维特单抗后，CADESI-03值和瘙痒率下降，这在单次使用2mg/kg剂量下最为明显。对瘙痒的影响持续时间为49天，CADESI-03为8周。另一项研究使用CADESI-03和瘙痒视觉模拟量表(PVAS)评估洛基维特单抗每月给药的有效性，治疗12周后，CADESI-03明显降低(从184降至57)。第一次给药后下降幅度最大(4周后CADESI-03从184降至76，下降54%)。这一发现与我们的研究相似，在CADESI-04，最大的下降幅度也是在4周后（从36.6到11.1；下降69%）。对于瘙痒，4周后观察到最大的下降，与我们研究中64% 的下降相比，下降大于51% 。

One study reported a gradual decrease in aTEWL (calculated as an average of 10 areas studied) that correlated with a decrease in the CADESI-03 in dogs with AD treated with ciclosporin. The decrease was found in the majority of areas assessed, although, similar to our study, not all decreases were shown to be statistically significant.TEWL values were reported to have been reduced in an evaluation of the efficacy of hydrocortisone acepionate; the clinical parameters CADESI and PVAS also decreased. Values for TEWL were used to evaluate treatment of cAD with oclacitinib; the results were inconclusive and for one body area the TEWL values were decreased, whereas increases were observed in other areas.

一项研究报告说，在用环孢素治疗特应性皮炎的犬中，aTEWL（计算10个研究区域的平均值）的量逐渐降低，与CADESI-03的降低有关。在大多数评估领域中发现了这一下降，但与我们的研究相似，并不是所有的下降都被证明具有统计学意义。在评价氢化可的松的疗效时，报告的TEWL值降低了，临床参数CADESI和PVAS 也降低了。TEWL值用于评估使用奥拉替尼治疗犬特应性皮炎的疗效；结果不确定，一个机体部位TEWL值下降，而其他部位的TEWL值增加。

The decrease in the average TEWL during lokivetmab treatment in our study may be explained by a reduction in inflammation in most of the examined areas. The reduction in inflammation led to normalization of epidermal barrier function. Self-trauma due to AD will damage the stratum corneum; the reduction of pruritus associated with lokivetmab treatment may help normalization of the skin barrier and lead to reduced TEWL values.

在我们的研究中，在洛基维特单抗治疗期间，aTEWL的下降可能是由于大多数被检查区域的炎症减少所致。 炎症的减少导致表皮屏障功能正常化。 由于特应性皮炎引起的自我损伤会损害角质层；与洛基维特单抗治疗相关的瘙痒的减少可能有助于皮肤屏障的正常化，并导致TEWL值的降低。

A limitation of our study is the choice of an pruritus assessment scale developed some time ago.We decided to use this scale to simplify the assessment of pruritus by the owners. The numerical pruritus scale contained six categories of pruritic behaviour similar to that used in the validated PVAS10 method.

我们研究的一个局限是选择了一个以前开发的瘙痒评估量表。我们决定用这个量表，简化宠主对瘙痒的评估。瘙痒评分表包含六类瘙痒行为，与已验证 PVAS10方法相似。

Lokivetmab administered at four week intervals was associated with decreases in the CADESI-04, pruritus and average TEWL values. When assessing response to treatment, it would seem reasonable to use the average TEWL calculated from several areas of the body given that a greater correlation is found in such cases. The assessment of individual parts of the body is much more variable and reduction in the TEWL value is not noted in every area during treatment.

每隔四周给药的洛基维特单抗与CADESI-04、瘙痒和aTEWL值的下降有关。在评估对治疗的反应时，使用从机体几个部位计算出的平均TEWL似乎是合理的，因为在这种情况下发现了更大的相关性。对机体各个部位的评估更加多变，在治疗过程中并没有注意到每个部位的TEWL值的下降。