Anogen的ELISA檢測試劑盒在細胞因子風(fēng)暴的檢測和治療中的應(yīng)用

COVID-19的爆發(fā)已經(jīng)奪去了全世界4000多人的生命，50000多名患者仍處于危急狀態(tài)。這種疾病的嚴(yán)重性，像許多其他急性呼吸道感染，如SARS和MERS，是由肺部病毒感染引起的促炎細胞因子水平升高引起的。

 

“細胞因子風(fēng)暴”是指免疫系統(tǒng)局部或全身產(chǎn)生過多的促炎細胞因子，以應(yīng)對多種感染性和非感染性刺激或物理和化學(xué)損傷。在嚴(yán)重急性呼吸綜合征(SARS)、H1N1和埃博拉病毒感染爆發(fā)期間，患者的高死亡率是細胞因子風(fēng)暴對身體功能產(chǎn)生負面影響的典型例子。目前，一旦細胞因子風(fēng)暴爆發(fā)，沒有抗素或抗病毒藥物能夠阻止它。

在Anogen，我們認(rèn)為應(yīng)對有害的細胞因子風(fēng)暴是一種新的生物防御策略。 自20世紀(jì)90年代以來，我們已開始細胞因子研究，并已開發(fā)出數(shù)百種雜交瘤，它們產(chǎn)生針對許多重要細胞因子的高特異性和高親和力單克隆抗體，包括腫瘤壞死因子-α、粒細胞巨噬細胞集落刺激因子、白介素-2、白介素-8、白介素-10、白介素-15、白介素-17、白介素-18和TGF- β。

此外，已經(jīng)開發(fā)了三種不同的用于檢測多種細胞因子表位的酶聯(lián)免疫試劑盒，并廣泛應(yīng)用于生物醫(yī)學(xué)研究。 

Multiplex Human Cytokine ELISA Kit (Inflammatory)

多重人細胞因子酶聯(lián)免疫試劑盒(炎性) 

Multiplex Human Cytokine ELISA Kit (M1/M2/MDSC Cytokines)

多重人細胞因子酶聯(lián)免疫試劑盒(M1/M2/MDSC細胞因子) 

Multiplex Human Cytokine ELISA Kit (Th1/Th2/Th17)

多重人細胞因子酶聯(lián)免疫試劑盒(Th1/Th2/Th17) 

在2003年非典爆發(fā)期間，Anogen與北京的醫(yī)院合作，發(fā)現(xiàn)患者體內(nèi)兩種促炎細胞因子IL-8(CXCL8)和MCP-1(MCAF /CCL2)顯著增加。

2004年，我們開發(fā)了抗趨化因子白細胞介素-8 (CXCL8)和單核細胞趨化蛋白-1 (CCL2)的人源化嵌合單克隆抗體，并在美國、歐洲、加拿大和中國獲得多項專利。臨床前研究中使用嵌合抗白介素-8抗體預(yù)防和治療急性呼吸窘迫綜合征的臨床前研究發(fā)表在《國際免疫藥理學(xué):針對趨化因子CXCL-8(白介素-8)的人源化單克隆抗體》上，可有效預(yù)防急性肺損傷。

如果你對其中任何一個感興趣，請不要猶豫與我聯(lián)系。我們期待您的來信。 

聯(lián)系單位：BioTNT上海冠泰生物科技有限公司（Anogen 中國總代理）

www.biotnt.com 

sales@biotnt.com 

我們正在尋求合作，進一步開發(fā)用于診斷和治療的細胞因子試劑盒和單克隆抗體。在由COVID-19感染期間致命的細胞因子風(fēng)暴引發(fā)的具有全部并發(fā)癥的疾病中，這些試劑盒和單克隆抗體應(yīng)該有利于挽救生命。

 

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白細胞介素-8是中性粒細胞的有效趨化因子、表皮細胞的生長因子和促血管生成因子。銀屑病是一種常見的皮膚病，其特征是鱗屑由快速生長的角質(zhì)細胞堆積而成，并伴有鱗屑周圍的炎癥和發(fā)紅。銀屑病組織中的白細胞介素-8水平高達150倍，表明其在發(fā)病機制中的作用。                 

Anogen-Yes生物技術(shù)公司是第一家利用抗白介素-8抗體治療銀屑病的公司。我們的專利技術(shù)(ABCREAM)是一種含有白介素-8中和抗體的外用軟膏。白介素-8中和抗體阻斷介導(dǎo)角質(zhì)形成細胞異常增殖和分化的白介素-8的活性，并增加病變附近的中性粒細胞浸潤以發(fā)揮抗炎作用。該軟膏在臨床試驗中被發(fā)現(xiàn)是有效的。49%至53.8%的患者在接受阿必利治療六周后，PASI評分改善超過60%，12.9%至15.3%的患者改善超過90%。

ABCream也可有效治療其他炎癥性皮膚病。初步觀察表明，它也可用于治療濕疹，一種常見的皮膚病。2001年獲中國食品藥品監(jiān)督管理局(CFDA)頒發(fā)的一類新藥證書。

手足口病(HFMD)是一種潛在的威脅生命的疾病，常見于大多數(shù)小于5歲的兒童。HFMD與腸道病毒71型(EV71)和柯薩奇病毒A6型(CV-A6)有關(guān)，屬于腸道病毒屬中的微小核糖核酸病毒科。盡管在包括歐洲和北美在內(nèi)的大多數(shù)國家都出現(xiàn)了全球性的疫情，但最大的HFMD疫情卻出現(xiàn)在亞太地區(qū)，原因不明。HFMD疫情不僅近年來有所增加，更嚴(yán)重的病例也更常見。更糟糕的是，HFMD病毒很可能會產(chǎn)生部分變異。重度HFMD的病死率較高。 高血糖癥、前白蛋白和白細胞增多癥目前被用于HFMD嚴(yán)重程度的臨床預(yù)測。然而，它們對HFMD患者既不敏感又無特異性。 已經(jīng)證明，由EV71或CV-A6感染的免疫細胞釋放的細胞因子和趨化因子有助于疾病的嚴(yán)重程度。不受控制的免疫反應(yīng)導(dǎo)致細胞因子表達異常升高，從而導(dǎo)致炎癥、組織損傷、肺水腫和其他病理損傷。這種現(xiàn)象被稱為“細胞因子風(fēng)暴”。 已經(jīng)證明，由EV71或CV-A6感染的免疫細胞釋放的細胞因子和趨化因子有助于疾病的嚴(yán)重程度。不受控制的免疫反應(yīng)導(dǎo)致細胞因子表達異常升高，導(dǎo)致炎癥、組織損傷、肺水腫和其他病理損傷。這種現(xiàn)象被稱為“細胞因子風(fēng)暴”。 最新研究表明，在多種細胞因子中，白細胞介素-2、白細胞介素-4、白細胞介素-6、白細胞介素-8、白細胞介素-10、單核細胞趨化蛋白-1、干擾素-γ、粒細胞集落刺激因子和腫瘤壞死因子-α在腸道病毒感染中增加。白細胞介素-8與HFMD病的嚴(yán)重程度密切相關(guān)，它與大量中性粒細胞浸潤組織和器官有關(guān)。盡管在HFMD患者中白細胞介素-6和白細胞介素-10升高，但與疾病的嚴(yán)重程度無關(guān)。因此，白細胞介素-8水平可作為嚴(yán)重腸道病毒感染早期的預(yù)測指標(biāo)。此外，人源化抗白介素-8抗體可預(yù)防致命并發(fā)癥，如腦干腦炎(BE)和肺水腫(PE)，以降低嚴(yán)重HFMD的死亡率。 我們的項目是開發(fā)一種新的用于早期預(yù)測嚴(yán)重HFMD病的多重細胞因子免疫測定(ELISA)的醫(yī)療裝置和一種有效的抗IL-8的治療性人源化抗體。

 

Cytokines are an integral part of the body’s immune system, but they also trigger adverse host responses such as fever, pain, and inflammation. “Cytokine storm” is a situation that the immune system produces excessive pro-inflammatory cytokines, locally or systemically, in response to a number of infectious and non-infectious stimulants, or physical and chemical damages. The high rate of patient death during the outbreak of severe acute respiratory syndrome (SARS), H1N1, and Ebola virus infection is a typical example of cytokine storm’s ferocity to negatively affect body functions. Currently, no antibiotics or antiviral drugs are able to stop the cytokine storm once it breaks out. At Anogen, we believe that tackling the harmful cytokine storm is a novel strategy of bio-defense.

We have initiated cytokine research since the 1990s and have developed hundreds of hybridomas that produce high specificity and high affinity monoclonal antibodies (mAbs) directed against many important cytokines, including TNF-α, GM-CSF, IL-2, IL-8, IL-10, IL-15, IL-17, IL-18, and TGF- β. In addition, three different ELISA kits for detection of multiple cytokine epitopes have been developed and widely applied to biomedical research.

Diseases with over production of cytokines:

 

 

IL-8 is a potent chemokine for neutrophils, growth factor of epidermal cells, and a pro-angiogenic factor. Psoriasis is a common skin disease characterized by scales that are built up with rapid growing keratinocytes accompanied by inflammation and redness around the scales. IL-8 levels elevate up to 150-fold in psoriatic tissue, suggesting its role in the pathogenesis.

Anogen-Yes Biotech is the first company utilizing anti IL-8 Ab to treat psoriasis. Our proprietary technology (ABCREAM) is a topical ointment containing IL-8 neutralization Ab. IL-8 neutralization Ab blocks the activity of IL-8 which mediates the abnormal proliferation and differentiation of keratinocytes, and increases neutrolphil infiltration in the lesion vicinity to exert the anti-inflammatory effect. The ointment was found to be effective during clinical trial. 49% to 53.8% of patients achieved a greater than 60% improvement and 12.9% to 15.3% of patients achieved a greater than 90% improvement in PASI scores after a six week treatment cycle with ABCream.

ABCream may be effective in treatment of other inflammatory skin conditions as well. Preliminary observations suggest that it may also be used for treatment of eczema, a common skin condition. It was awarded Class I New Drug Certificate issued by China Food and Drug Administration (CFDA) in 2001. 

HFMD and IL-8

Hand, foot and mouth disease (HFMD) is a potentially life-threatening illness commmonly seen in children mostly younger than five years of age. HFMD associated with Enterovirus 71 (EV71) and Coxsackievirus A6 (CV-A6), members of the Picornaviridae family in the genus Enterovirus.  Although present globally in most countries including Europe and North America, the largest HFMD outbreaks has been seen in the Asia-Pacific area, for unknown reasons. HFMD outbreaks are not only increased in recent years, but more severe cases are also more common. Even worse, the HFMD virus is likely to develop partial variations. The fatality rate of severe HFMD is higher.

Hyperglycemia, Pre-albumin and leukocytosis are currently used for clinical prediction of the severity of HFMD. However, they are both insensitive and nonspecific in HFMD patients.

It has been proved that cytokines and chemokines released by EV71 or CV-A6 infected immune cells contribute to the disease severity. The uncontrolled immune responses result in abnormally elevated expression of cytokines that cause inflammation, tissue damage, pulmonary edema and other pathological damage. This phenomenon is known as "Cytokine Storm".

It has been proved that cytokines and chemokines released by EV71 or CV-A6 infected immune cells contribute to the disease severity. The uncontrolled immune responses result in abnormally elevated expression of cytokines that cause inflammation, tissue damage, pulmonary edema and other pathological damage. This phenomenon is known as "Cytokine Storm".

The latest research showed that among many cytokines, IL-2, IL-4, IL-6, IL-8, IL-10, MCP-1, IFN-γ, GM-CSF and TNF-α increase in enterovirus infection. IL-8 is strongly associated with the severity of HFMD with massive neutrophil infiltration of tissue and organ. IL-6 and IL-10, although elevated in HFMD patients, are not related to the disease severity. Thus, IL-8 level could be used as a predictive marker in the early stages of severe enterovirus infection. In addition, humanized anti-IL-8 antibody may prevent fatal complications such as brainstem encephalitis (BE), and pulmonary edema (PE) to reduce the mortality of severe HFMD.

Our project is to develop a new medical device of multiplex cytokine immuno-assay (ELISA) for early prediction of severe HFMD and an effective therapeutic humanized antibody against IL-8.

 

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