CN117050177B - 血液分离的免疫细胞联合药物治疗癌症的用途 - Google Patents
血液分离的免疫细胞联合药物治疗癌症的用途 Download PDFInfo
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Abstract
本发明涉及血液分离的免疫细胞联合药物治疗癌症的用途。本发明针对宫颈癌，开发了特异性的DC疫苗。同时，针对IL‑8开发了特异性的单克隆抗体，所述单克隆抗体单独或者联合DC疫苗使用后，可以有效的抑制宫颈癌瘤重的增加，能够有效的实现宫颈癌的治疗，可以用于制备抗肿瘤药物，应用价值较高。
Description
技术领域
本申请涉及生物领域，具体的涉及血液分离的免疫细胞联合药物治疗癌症的用途。
背景技术
HPV是宫颈上皮内瘤变(CIN)和宫颈癌的重要病因。鳞状细胞癌是宫颈癌最常见的组织学类型，其他常见的组织学类型还包括宫颈腺癌和腺鳞癌。硏究表明年轻成年女性中HPV感染的患病率高达40％-80％，而宫颈癌是未解决的HPV感染的罕见终末期。在流行病学方面,至少有12种HPV被归类为致癌的高危(HR)类型(HPV16/18/31/35/39/45/51/52/56/58/66/68)，上述HPV16和HPV18是最常出现的HR-HPV类型，并引起约70％的宫颈癌。HPV是具有221种鉴定类型的小型非包膜双链DNA病毒,这些病毒具有8kb的基因组，可编码早期调节蛋白(E1、E2、E4、E5、E6和E7)和晚期结构蛋白(L1和L2)oE1和E2蛋白调节HPVDNA的复制和转录；E4通过破坏上皮细胞角蛋白来协助病毒释放；E5负责诱导宿主细胞中的生长因子；E6结合和降解肿瘤抑制基因p53和促凋亡蛋白BAK；E7结合并抑制肿瘤抑制因子pRb。
基于DC的疫苗基于DC的HPV疫苗已成为对抗HPV相关恶性肿瘤的潜在治疗疫苗，因为它们不仅是主要的抗原呈递细胞，也可以作为天然佐剂增强针对癌症的抗原特异性免疫疗法的效力。目前尽管可以通过用针对促凋亡分子的小干扰RNA(siRNA)转染DC，但仍有因为技术要求导致无法大规模生产、有效的疫苗给药途径尚不确定需要患者提供足够的自体DC、转导效率低、终末分化DC无法体外扩增以及DC寿命有限等局限。基于DC的疫苗可以分为用HPV特异性肽/蛋白质抗原脉冲化的DC或用编码外来抗原的DNA/病毒载体转导的DC，同样因为未成熟的DC缺乏完整的T细胞共刺激活性，导致大多数试验必须使用成熟的DC。有学者先后将DC疫苗用于治疗早期宫颈癌和复发、难治性宫颈癌患者，结果均提示无临床获益，仅部分患者有血清学效应。但研究发现甘草水提取物(GUWE)可以促进DC的成熟并增加了机体内细胞因子的产生，同时证明GUWE诱导了HPV特异性细胞应答并抑制了荷瘤小鼠的肿瘤生长，这在某种程度上为DC疫苗的开发提供了一种策略。
肿瘤免疫治疗是一种依靠免疫系统识别和直接杀死肿瘤细胞，人为地增强机体的免疫功能以达到治疗疾病目的的治疗方法。以免疫检查点抑制剂为代表的免疫治疗为改善宫颈癌治疗和生存带来了希望。在正常、非肿瘤侵袭的情况下，免疫检查点分子可维持自身耐受、预防自身免疫并保护正常细胞免受免疫攻击或感染期间的长期炎症。在此稳态期间，细胞免疫受激活信号(如共刺激分子，抗原提呈细胞所表达的)和抑制信号(如免疫检查点)的调节。随着组织癌变，肿瘤细胞会进行免疫编辑，通过上调免疫检查点分子逃避免疫监视，从而能够不受控制进行增长和扩散，并产生免疫抑制性肿瘤微环境(TME)，同时下调效应T细胞的抗肿瘤活性。免疫检查点抑制剂(ICIs)是目前国际上肿瘤免疫治疗的主要研究和应用方向，其主要是解除肿瘤对免疫的耐受/屏蔽作用，让免疫细胞重新认识肿瘤细胞，对肿瘤产生攻击，通过抑制或共刺激信号等一系列途径以调节T细胞活性来杀伤肿瘤细胞的治疗方法。
目前研究和应用的免疫检查点疗法集中在靶向PD-1/PD-L1(程序性死亡受体-1/程序性死亡配体-1)和CTLA-4(细胞毒性T淋巴细胞相关蛋白4)的拮抗性抗体。PD-1/PD-L1是主要的免疫检查点机制。PD-1表达在效应免疫细胞表面，可结合TME中肿瘤细胞或免疫抑制细胞表达的PD-L1，“免疫逃逸”最主要的途径就是通过PD-1/PD-L1通路实现的。另一个重要的免疫检查点是CTLA-4，CTLA-4在T细胞表面表达，一旦与其在APC上发现的配体CD80或CD86结合，就会下调T细胞的活化和细胞毒性；此外，CTLA-4可以通过在调节性T细胞(Treg)上的表达来调节T细胞的内在和外在活化。该免疫检查点通路在自身免疫性疾病中也起着重要作用。PD-1和CTLA-4免疫检查点的阻断可以通过单克隆抗体(mAb)的拮抗剂进行调节，分别用于增强T细胞活化和消除对T细胞毒性的抑制，以重新激活T细胞以攻击肿瘤。相比较而言，阻断PD-1/PD-L1主要通过消除CD8+T细胞耗竭发挥作用；而阻断CTLA-4通路主要通过激活新的T细胞和抑制Treg细胞发挥作用，在免疫抑制性TME中，ICIs对CTLA-4的抑制有助于逆转免疫系统抑制并促进肿瘤清除。基于KEYNOTE-158临床试验，FDA已批准PD-1抑制性抗体帕博利珠单抗用于宫颈癌治疗。在KEYNOTE-158试验中，98名先前接受过治疗的晚期宫颈癌患者接受每3周200mg帕博利珠单抗的治疗，直至疾病进展、不可耐受的毒性或撤回知情同意，治疗周期最长为2年，试验结果显示客观缓解率(ORR)为12.2％，完全缓解或部分缓解的患者都呈PD-L1阳性；对于至少接受过一次化疗的患者亚组，ORR上升到14％；先前接受过化疗患者的ORR提升表明了免疫治疗与其他肿瘤治疗手段相结合的潜力。一项正在进行的转移性/复发性宫颈癌患者的I/II期临床试验表明，CTLA-4抑制剂伊匹单抗具有良好的耐受性，并能促进免疫激活。其他临床试验研究了不同ICIs在宫颈癌中的应用。目前ICIs治疗也遇到了瓶颈——PD-1单抗单药用于复发或转移性宫颈癌二线及以上治疗有效率较低。双免疫联合疗法及双特异性单克隆抗体等新型抗体药物，在宫颈癌领域近年来也有突破性进展。Checkmate-358研究探索了帕博利珠单抗联合伊匹单抗用于复发/转移性宫颈癌的治疗，结果显示二线及以上治疗的ORR达到了36.4％(Nivo1+Ipi3)，但两药联合的安全性问题不容忽视。临床迫切需要寻找效毒比更高的组合方案或新型药物。
发明内容
本发明克服现有技术的不足，提供了一种单克隆抗体联合DC细胞一起治疗宫颈癌的技术方案，所述方案具有较好的治疗效果。
一方面，本发明提供一种特异性针对IL-8的单克隆抗体。
所述的IL-8单克隆抗体命名为3E8-2，其轻链可变区序列如SEQ ID NO：1所示，其重链可变区序列如SEQ ID NO：2所示。
所述抗体具有较强的特异性以及亲和活力。具有抑制宫颈癌细胞的效果。
进一步的，本发明提供了IL-8单克隆抗体3E8-2在制备用于治疗宫颈癌的药物组合物中的用途。
具体的，所述的单克隆抗体可以是如单克隆抗体或其片段；嵌合单克隆抗体(如人-鼠嵌合单克隆抗体)；完全人单克隆抗体；重组人单克隆抗体；人源化抗体片段。任选地，IL-8单克隆抗体是单克隆抗体的功能片段或包含功能片段的融合蛋白，如Fab、F(ab')2、Fv，并且优选为Fab。优选地，片段被聚乙二醇化或包封(例如，为了稳定性和/或缓释)。
如本文所讨论，认为抗体或免疫球蛋白分子的氨基酸序列的变化涵盖于本发明中，条件是氨基酸序列的变化保持至少75％，更优选至少80％、90％、95％和最优选99％。包括其间的某些百分比，例如75％、76％、77％、78％、79％、80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％和99％序列同一性。尤其，包括保守性氨基酸置换。保守性置换是在与侧链相关的氨基酸家族中发生的置换。遗传编码的氨基酸通常分为以下家族：(1)酸性氨基酸为天冬氨酸、谷氨酸；(2)碱性氨基酸为赖氨酸、精氨酸、组氨酸；(3)非极性氨基酸为丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸、色氨酸，和(4)不带电的极性氨基酸为甘氨酸、天冬酰胺、谷氨酰胺、半胱氨酸、丝氨酸、苏氨酸、酪氨酸。亲水性氨基酸包括：精氨酸、天冬酰胺、天冬氨酸、谷氨酰胺、谷氨酸、组氨酸、赖氨酸、丝氨酸和苏氨酸。疏水性氨基酸包括丙氨酸、半胱氨酸、异亮氨酸、亮氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、色氨酸、酪氨酸和缬氨酸。其它氨基酸家族包括(i)为脂肪族羟基家族的丝氨酸和苏氨酸；(ii)为含有酰胺的家族的天冬酰胺和谷氨酰胺；(iii)为脂肪族的丙氨酸、缬氨酸、亮氨酸和异亮氨酸；和(iv)为芳香族的苯丙氨酸、色氨酸和酪氨酸。例如，有理由期望用异亮氨酸或缬氨酸单独置换亮氨酸，用谷氨酸单独置换天冬氨酸，用丝氨酸单独置换苏氨酸，或用结构相关的氨基酸对氨基酸进行相似置换对所得分子的结合或性质无较大影响，特别是如果置换不牵涉骨架位点内的氨基酸时。通过检测多肽衍生物的特异性活性可易于确定氨基酸变化是否产生功能性肽。本文对检测进行了详细描述。本领域的普通技术人员可易于制备抗体或免疫球蛋白分子的片段或类似物。片段或类似物的优选氨基端和羧基端出现在功能结构域边界附近。可通过比较核苷酸和/或氨基酸序列数据和公共或私有序列数据库鉴定结构和功能结构域。
更进一步的，本发明提供了IL-8单克隆抗体3E8-2联合DC疫苗在制备用于治疗宫颈癌的药物组合物中的用途。其中DC疫苗是将宫颈癌细胞通过反复冻融后获得肿瘤细胞冻融裂解物后刺激DC细胞后制备获得的。
更进一步的，所述的DC疫苗可以采用本领域较为成熟的其他制备方法来制备获得。
更进一步的，本发明的单克隆抗体还可以负载在DC疫苗上来发挥作用，也可以分别给药的形式来发挥作用。
组合物可能以单剂量给药，或者多剂量给药。在多剂量的计划中，接种的主要过程可以包括如1-10个独立的剂量，随后在间隔的时间给药其它剂量，从而保持或者增强免疫应答，例如在第1个月进行第二剂量，并且如果需要，在几个月后给药随后的剂量(或者多个剂量)。
本发明的疫苗可以以一个或多个“单位剂量”的形式来提供，这取决于是否使用核酸载体、是否使用最终纯化的蛋白或最终的疫苗形式。单位剂量定义为含有预先确定量的治疗组合物，计算所述的治疗组合物以在其给药的时候，即在期待的途径下和治疗区域中产生期待的应答。要给药的量，以及具体的途径和制剂是临床技术领域技术人员所了解的。也可以对要治疗的受体进行评估，尤其对于受体的免疫系统的状态和所期待的保护进行评估。
有益效果
本发明针对宫颈癌，开发了特异性的DC疫苗。同时，针对IL-8开发了特异性的单克隆抗体，所述单克隆抗体单独或者联合DC疫苗使用后，可以有效的抑制宫颈癌瘤重的增加，能够有效的实现宫颈癌的治疗，可以用于制备抗肿瘤药物，应用价值较高。
附图说明
图1IL-8单克隆抗体3E8-2的特异性鉴定结果图
图2各组肿瘤重量结果图
具体实施方式
通过以下实施例进一步说明本公开文本，所述实施例不应解释在范围或精神上将本公开文本限制在本文所述的具体程序内。应当理解，提供这些实施例是为了说明某些实施方案，并且并非意在借此限制本公开文本的范围。应进一步理解，在不背离本公开文本的精神和/或所附权利要求书的范围的情况下，可能不得不诉诸本领域技术人员可能提出的各种其他实施方案、修改及其等同物。
实施例1DC疫苗的制备
常规培养宫颈癌CaSki细胞到对数期，PBS洗涤3次，经液氮迅速冷冻后，立即放入37℃水浴中至刚刚融化，即再次放入液氮迅速冷冻，反复5次冻融循环获得肿瘤细胞冻融裂解物，10000r/min低温离心20min，上清用0.22μm针头滤器过滤，Lowry法测蛋白含量，-20℃保存。
在无菌条件下，从KM小鼠股骨和胫骨冲洗骨髓腔收集骨髓细胞，过200目网，1000r/min离心5min，弃上清，以Tris-NH4Cl处理2～3min去除红细胞，用PBS洗涤2次，RPMI-1640培养液洗1次后，将细胞悬于含rmGM-CSF(100μg/L)、rmIL-4(100μg/L)、rmIL-2(30μg/L)的含10％胎牛血清的RPMI-1640完全培养液中(细胞浓度为1×109cells/L)培养。每2d半量换液加前述全量细胞因子1次，第6d时，用100mg/L CaSki细胞冻融抗原刺激DCs 24h，收集疏松贴壁细胞，流式细胞术检测细胞均高表达CD11c(86.53％±5.34％)、CD80(89.31％±3.62％)和CD86(92.57％±4.18％)。
实施例2CCK-8检测DC疫苗的活性
拉颈处死KM小鼠，在无菌条件下取出脾脏，研磨成单细胞，经200目细胞筛过滤，收集细胞并加入小鼠淋巴细胞分离液4mL，800×g离心30min，取淋巴细胞层，用含10％胎牛血清的RPMI1640完全培养液洗涤2次，重悬并计数细胞。取负载CaSki细胞裂解物的DCs疫苗及未处理的DCs，分别与淋巴细胞以1:10的比例混合共培养24h，诱导产生CTLs作为效应细胞。
取上述效应细胞与作为靶细胞的CaSki细胞混合，效靶比例分别为20:1和10:1，接种于96孔板中，以完全培养液作为空白对照，CaSki细胞为对照组，每组设3个复孔。细胞培养24h后，按CCK-8检测试剂盒说明书的方法测定细胞毒效应，在酶联免疫检测仪上测定波长为570nm处各孔的吸光度(D)值，按以下公式计算细胞杀伤效率。细胞杀伤效率＝[1-(实验组D值﹣空白对照组D值)/(对照组D值-空白对照组D值)]×100％。结果如表1所示。
表1CCK-8法检测CTLs对CaSki细胞的杀伤率
(*P<0.05)
从表1可以看出，负载CaSki细胞裂解物的DCs疫苗相对于未处理的DCs具有更好的诱导CTLs显著的杀伤CaSki细胞的效果。
实施例3IL-8单克隆抗体的制备
白介素8(IL8)重组蛋白，LMAI Bio，货号：ICA080Bo01。
以重组蛋白IL8为抗原，免疫8周龄雌性BALB/c小鼠。首次免疫，抗原与等体积的弗氏完全佐剂混合乳化，颈背部皮内多点注射，50μg/只。3周后进行二免，抗原与等体积弗氏不完全佐剂混合乳化，颈背部皮下多点注射，50μg/只。3周后进行三免，免疫剂量与途径同二免。三免1周后，小鼠断尾采血分离血清，以重组蛋白IL8所包被的酶标反应板用间接ELISA的方法测定血清抗体效价，其中效价最高达到了1:64000，该小鼠达到融合以制备单克隆抗体的标准。在细胞融合前3d，进行加强免疫，小鼠腹腔注射抗原，60μg/只。
融合前先将lmL的PEG-1500和20mL的RPMI-1640基础培养基于37℃预温。取小鼠的脾细胞与骨髓瘤细胞按10：1比例混合于50mL离心管内，1000r/min离心5min。离心后弃上清。然后轻轻敲击离心管底使细胞松散。在37℃水浴中沿管壁加入PEG 1mL，60s内加完，边加边转动离心管，不宜吹打，加完后在37℃水浴中静置反应1min，再加入RPMI-1640基础培养基以终止PEG的作用，在1min内加完1mL，再在1min内加完2mL，然后逐渐加快速度，在5min内加完15mL。加终止液时应缓慢加入，动作应轻柔，边加边轻轻搅拌。不宜吹打以免使刚融合的细胞分离。融合细胞终止后于800r/min离心6min，并吸干以防残留PEG。再加入预温的HAT培养基至所需的细胞浓度，并轻轻将细胞混匀，然后100μL/孔滴板。细胞融合7d后，用间接ELISA法检测抗体，选择抗体分泌阳性孔，用HT培养基扩大培养后，采用有限稀释法接种于96孔培养板内，选择单个细胞集落孔的培养上清作抗体检测，阳性者用同样方法再次克隆，直至克隆后有杂交瘤细胞生长的抗体分泌阳性率为100％。经过4次亚克隆后，得到1株阳性反应最强最稳定分泌抗IL-8蛋白的杂交瘤细胞株，命名为3E8-2。将杂交瘤细胞注射到小鼠腹腔，收获腹水后利用正辛酸饱和硫酸铵法纯化，获得纯度较高的3E8-2单克隆抗体，调整抗体浓度为1mg/mL备用。
实施例4IL-8单克隆抗体3E8-2的特异性鉴定
用IL-8重组蛋白、His标签蛋白做常规的SDS-PAGE分析。电泳结束后，采用半干转印法，将蛋白转印到NC膜上；将NC膜放在含5mg/L脱脂乳的PBS封闭液中过夜，用含5mg/L吐温-20的PBST洗涤液洗膜3次，10min/次；然后将3E8-2单抗做1:4000稀释，与NC膜在37℃作用2h，用PBST洗膜5次，10min/次；再与1:4000稀释的山羊抗鼠酶标二抗在室温下孵育1h，用PBST彻底洗涤，加入DAB底物显色液显色，当出现明显条带时终止反应，观察结果。结果如图1所示。
从图1可以看出，IL-8单克隆抗体3E8-2能够特异性的结合IL-8，而不与His蛋白结合，表现出较好的特异性。
实施例5IL-8单克隆抗体3E8-2亲和力的鉴定以及序列鉴定
利用间接ELISA方法测定单克隆抗体的亲和力，以1g/mL重组蛋白IL8包被酶标反应板，封闭后加入倍比稀释的单克隆抗体，以HRP标记的山羊抗小鼠IgG为二抗，酶标仪读取OD450吸光值。当连续几个稀释度的OD450读数不再增大时视为抗原抗体100％结合，以抗体浓度(mol/L)为横坐标，OD450吸光值为纵坐标做散点图，生成对数趋势线和公式。将OD450最大值的一半带入公式，求出此时的抗体浓度即为单克隆抗体的亲和力解离常数(Kd)。结果如表2所示。
表2IL-8单克隆抗体3E8-2亲和力结果
| 抗体名称 | Kd |
| 3E8-2 | 2.53×10-9 |
利用间接ELISA方法测定单克隆抗体的亲和力，结果表明IL-8单克隆抗体3E8-2的亲和力解离常数(Kd)为2.53×10-9，亲和力较强。
将IL-8单克隆抗体3E8-2的杂交瘤细胞进行抗体轻重链可变区序列鉴定，委托上海生工完成，测序得到IL-8单克隆抗体3E8-2的轻链可变区序列如SEQ ID NO：1所示，其重链可变区序列如SEQ ID NO：2所示。
实施例6单抗单独及联合给药实验
Balb/c(nu/nu)均为雌性，体质量16～18g，SPF级，购买于北京实验动物研究中心。将CaSki细胞悬液调整为3×107/ml。在裸鼠右侧腋下靠背侧皮下接种CaSki细胞悬液0.2ml。
实验裸鼠用随机数字表法分为6组，每组各10只。
空白对照组(1组)注射等体积PBS；
阴性对照组(2组)注射等体积IgG；
治疗组(3组)注射抗IL-8抗体，100μg/次，每3d 1次；
治疗组(4组)注射抗IL-8抗体，500μg/次，每3d 1次；
DCs疫苗组(5组)注射实施例1的DCs1×106个(0.1mL)，每5d 1次；
DCs疫苗联合单抗治疗组(6组)注射实施例1的DCs 1×106个(0.1mL)，每5d 1次；注射抗IL-8抗体，100μg/次，每3d 1次；
阳性对照组(7组)注射市售抗IL-8抗体(大连亚维药业有限公司)，100μg/次，每3d1次；
以上各组自肿瘤接种后4d起开始注射，均为腹腔内注射，共4周。每组均先各组眼球取1ml血液用于检测血清中IL-8蛋白，随后将小鼠处死后，取瘤体测量肿瘤重量，结果如图2所示。
从图2可以看出，治疗组(3-7组)肿瘤重量小于对照组(1-2组)，差异有统计学意义(均P＜0.05)；1组与2组间差异无统计学意义。从3组和4组的结果可以看出，随着抗IL-8抗体剂量的增加，肿瘤体积也随之减小，不同剂量组间比较差异有统计学意义。在同浓度的单抗处理条件下，本发明的单克隆抗体的治疗效果要比市售的单抗效果要好，第3组的瘤重明显比第7组的瘤重要小。此外，第5组单独使用DC疫苗治疗后，也能显著的抑制瘤重，特别是将DC疫苗和单抗抑制使用后，抑制移植瘤生长效果明显，说明将单抗和DC疫苗联用后能够协同的抑制宫颈癌的生长。裸鼠经抗IL-8抗体治疗后，均健康活泼，饮食正常，生长状态良好。
将前述各组眼球取血放置2h后，离心半径9.2cm，3000r/min离心5min，收集上层血清。此后严格按照ELISA试剂盒说明书同批进行检测。用自动化酶联免疫分析仪测定450nm处的A值，以A值为Y轴，相应试剂盒中IL-8标准品的浓度为X轴做标准曲线。测定各组裸鼠血清中IL-8浓度。结果如表3所示。
表3各组小鼠血清中IL-8蛋白表达结果(pg/ml)
| 组别 | 表达量 |
| 1组 | 408.2±53.7 |
| 2组 | 400.4±48.6 |
| 3组 | 200.1±38.4 |
| 4组 | 95.3±17.5 |
| 5组 | 372.3±54.1 |
| 6组 | 86.4±10.7 |
| 7组 | 290.6±37.8 |
从表3的结果可以看出，3-4、6-7的治疗组与1-2的对照组相比IL-8蛋白表达显著降低(均P<0.05)，且有剂量依赖性，4组比3组抑制效果更显著；两对照组1-2组之间差异无统计学意义。者说明IL-8抗体能够有效的抑制IL-8蛋白的表达量。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，但本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (8)
1.一种IL-8单克隆抗体3E8-2，其特征在于所述抗体的轻链可变区序列如SEQ ID NO：1所示，其重链可变区序列如SEQ ID NO：2所示。
2.如权利要求1所述的单克隆抗体在用于制备治疗宫颈癌的药物组合物中的用途。
3.如权利要求2所述的用途，其特征在于所述的药物组合物还含有药学上可接受的载体。
4.如权利要求3所述的用途，其特征在于所述药物组合物是注射剂的形式。
5.如权利要求1所述的单克隆抗体在制备用于特异性检测IL-8的试剂盒中的用途。
6.一种血液分离的免疫细胞联合IL-8单克隆抗体在制备用于治疗宫颈癌的药物组合物中的用途；其中所述的血液分离的免疫细胞是DC疫苗，该DC疫苗是将宫颈癌细胞通过反复冻融后获得肿瘤细胞冻融裂解物后刺激DC细胞后制备获得的；所述的IL-8单克隆抗体为3E8-2，其轻链可变区序列如SEQ ID NO：1所示，其重链可变区序列如SEQ ID NO：2所示。
7.如权利要求6所述的用途，其特征在于所述的药物组合物还含有药学上可接受的载体。
8.如权利要求7所述的用途，其特征在于所述药物组合物是注射剂的形式。
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| WO2016168264A1 (en) * | 2015-04-13 | 2016-10-20 | Kiromic, Llc | Methods and compositions for treating cancer with dendritic cells |
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| WO2016168264A1 (en) * | 2015-04-13 | 2016-10-20 | Kiromic, Llc | Methods and compositions for treating cancer with dendritic cells |
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| 抗白细胞介素8单克隆抗体对子宫颈癌的抑制作用;张艳丽;吴素慧;李雪;高艺敏;孙静汾;尚海霞;杨彦林;肿瘤研究与临床;第30卷(第003期);第145-151, 156页 * |
| 白细胞介素-8与肿瘤的研究进展;杨蒙;刘晓斌;;中国医药导报(第26期);第39-43+53页 * |
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