公开(公告)号：US20150252420A1

公开(公告)日：2015-09-10

申请号：US14642546

申请日：2015-03-09

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: C12Q1/68 ,  C07K16/28 ,  G01N33/569

CPC classification number: C12Q1/6881 ,  C07K16/28 ,  C07K2317/34 ,  C12N5/0087 ,  C12N5/0638 ,  G01N33/56972 ,  G01N2333/295 ,  G01N2333/5437 ,  G01N2333/70517

Abstract: Methods are provided for isolating CD8+ T cells. Furthermore, methods for purifying a subset of IL-13 expressing CD8 T cells are provided, such methods comprising the steps of marking the CD8+ T cells by labeling CD8, or selectively removing non-CD8 cells, and then purifying a subset of IL-13 expressing CD8+ T cells by marking a human biomarker such as C10orf128. Related antibodies and antiserums are also described, such antibodies related to a cell surface domain peptide for biomarker C10orf128, and human homologs of related mouse “activated” CD8IL-13 cell surface biomarkers Tm4sf19 and 1830127L07Rik.

Abstract translation: 提供了隔离CD8 + T细胞的方法。 此外，提供了用于纯化表达IL-13的CD8 T细胞亚群的方法，所述方法包括以下步骤：通过标记CD8或选择性除去非CD8细胞来标记CD8 + T细胞，然后纯化IL-13的子集 通过标记人类生物标志物如C10orf128表达CD8 + T细胞。 还描述了相关抗体和抗血清，与生物标记C10orf128的细胞表面结构域肽相关的抗体以及相关小鼠“活化的”CD8IL-13细胞表面生物标志物Tm4sf19和1830127L07Rik的人类同源物。

公开(公告)号：US09683262B2

公开(公告)日：2017-06-20

申请号：US14642546

申请日：2015-03-09

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: C12Q1/00 ,  C12Q1/68 ,  C12N5/07 ,  C12N5/16 ,  G01N33/569 ,  C07K16/28 ,  C12N5/00 ,  C12N5/0783

CPC classification number: C12Q1/6881 ,  C07K16/28 ,  C07K2317/34 ,  C12N5/0087 ,  C12N5/0638 ,  G01N33/56972 ,  G01N2333/295 ,  G01N2333/5437 ,  G01N2333/70517

Abstract: Methods are provided for isolating CD8+ T cells. Furthermore, methods for purifying a subset of IL-13 expressing CD8 T cells are provided, such methods comprising the steps of marking the CD8+ T cells by labeling CD8, or selectively removing non-CD8 cells, and then purifying a subset of IL-13 expressing CD8+ T cells by marking a human biomarker such as C10orf128. Related antibodies and antiserums are also described, such antibodies related to a cell surface domain peptide for biomarker C10orf128, and human homologs of related mouse “activated” CD8IL-13 cell surface biomarkers Tm4sf19 and 1830127L07Rik.

公开(公告)号：US20140274783A1

公开(公告)日：2014-09-18

申请号：US14215144

申请日：2014-03-17

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: C12N5/0783 ,  G01N33/569

CPC classification number: G01N33/56977 ,  C12N5/0087 ,  C12N5/0638 ,  G01N33/56972 ,  G01N2333/295 ,  G01N2333/70517

Abstract: Isolated, Chlamydia-specific, IL-13 expressing CD8+ T cell clones are provided for understanding the biology of Chlamydia infection and screening of therapeutics. Furthermore, methods are provided for isolating CD8+ T cells comprising the steps of infecting or identifying a naturally infected mammal with at least one species of Chlamydia causing bacteria, allowing the bacteria to clear or for a T cell immune response to the natural infection, collecting immune splenocytes from the mammal, providing at least one antigen from a Chlamydia-specific bacteria, and expanding and depleting the CD4+ T cell population or purifying the CD8 T cell population to isolate IL-13 expressing CD8+ T cell clones.

Abstract translation: 提供分离的衣原体特异性IL-13表达CD8 + T细胞克隆以了解衣原体感染的生物学和筛选治疗剂。 此外，提供了用于分离CD8 + T细胞的方法，包括以下步骤：用至少一种引起衣原体的细菌感染或鉴定天然感染的哺乳动物，允许细菌清除或对天然感染的T细胞免疫应答，收集免疫 来自哺乳动物的脾细胞，提供来自衣原体特异性细菌的至少一种抗原，以及扩增和消耗CD4 + T细胞群体或纯化CD8T细胞群以分离表达IL-13的CD8 + T细胞克隆。

公开(公告)号：US10663467B2

公开(公告)日：2020-05-26

申请号：US15627755

申请日：2017-06-20

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: G01N33/569 ,  G01N33/68 ,  G01N33/50 ,  C07K16/28 ,  C12Q1/6881 ,  C12N5/0783 ,  C12N5/00 ,  C12Q1/6883

Abstract: Methods for identifying and isolating CD8 T cells that produce interleukin-13 upon activation are provided. The present methods leverage one or more newly-identified biomarkers to identify such CD8 T cells and, in certain cases, sort the same. Certain methods comprise obtaining a sample from a mammal, quantifying a level of expression of one or more biomarkers therein, and determining if the level of expression is elevated as compared, wherein an elevated expression level is indicative of an active disease state. Antisera and antibodies are also provided. In particular, an anti-C10orf128 antiserum formulated against a particular peptide is provided, such anti-C10orf128 antiserum characterized in that it identifies a subset of CD8 T cells that produce interleukin-13 upon activation.

公开(公告)号：US20170363626A1

公开(公告)日：2017-12-21

申请号：US15627755

申请日：2017-06-20

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: G01N33/569 ,  G01N33/68 ,  C07K16/28 ,  G01N33/50 ,  C12Q1/68

CPC classification number: G01N33/56972 ,  C07K16/28 ,  C07K2317/24 ,  C07K2317/34 ,  C07K2317/76 ,  C12N5/0087 ,  C12N5/0638 ,  C12Q1/6881 ,  C12Q1/6883 ,  C12Q2600/158 ,  G01N33/505 ,  G01N33/6869 ,  G01N2333/295 ,  G01N2333/5409 ,  G01N2333/5437 ,  G01N2333/70517 ,  G01N2800/26

Abstract: Methods for identifying and isolating CD8 T cells that produce interleukin-13 upon activation are provided. The present methods leverage one or more newly-identified biomarkers to identify such CD8 T cells and, in certain cases, sort the same. Certain methods comprise obtaining a sample from a mammal, quantifying a level of expression of one or more biomarkers therein, and determining if the level of expression is elevated as compared, wherein an elevated expression level is indicative of an active disease state. Antisera and antibodies are also provided. In particular, an anti-C10orf128 antiserum formulated against a particular peptide is provided, such anti-C10orf128 antiserum characterized in that it identifies a subset of CD8 T cells that produce interleukin-13 upon activation.

公开(公告)号：US20170002419A1

公开(公告)日：2017-01-05

申请号：US15218141

申请日：2016-07-25

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: C12Q1/68 ,  G01N33/577 ,  G01N33/564

CPC classification number: C12Q1/6883 ,  C12Q2600/106 ,  C12Q2600/118 ,  C12Q2600/158 ,  G01N33/5047 ,  G01N33/564 ,  G01N33/577 ,  G01N33/6893 ,  G01N2333/47 ,  G01N2800/102 ,  G01N2800/24 ,  G01N2800/245 ,  G01N2800/52

Abstract: A previously unknown T cell receptor (TCR) activation pathway dependent on the aryl hydrocarbon receptor conferring resistance to calcineurin inhibitors and mTOR inhibitors is disclosed, including application of this pathway to the diagnosis and treatment of certain disease states refractory to treatment with calcineurin inhibitors. This alternative TCR activation pathway uniquely exists in a subset of CD8 T cells expanded in the setting of chronic rejection or rheumatoid arthritis. Expansion of this newly discovered calcineurin and mTOR inhibitor resistant CD8 T cell subset in humans can be quantified by measuring levels of certain biomarkers in the circulating CD8 T cell pool, such as Pla2g4a, to diagnose disease states mediated thereby. Additionally, methods for diagnosing ongoing active inflammation mediated by this resistant CD8 T cell subset in either chronic rejection or rheumatoid arthritis are provided, which comprise measuring levels of the biomarker Scin in the circulating CD8 T cell pool.

Abstract translation: 公开了一种先前未知的依赖于赋予对钙调神经磷酸酶抑制剂和mTOR抑制剂的抗性的芳基烃受体的T细胞受体（TCR）激活途径，包括将该途径应用于诊断和治疗用钙调神经磷酸酶抑制剂治疗的某些疾病状态。 该替代的TCR激活途径独特存在于在慢性排斥反应或类风湿性关节炎的环境中扩增的CD8T细胞的亚群中。 通过测量循环CD8 T细胞池（如Pla2g4a）中某些生物标志物的水平，可以量化这种新发现的钙调神经磷酸酶和mTOR抑制剂抗性CD8 T细胞亚群在人中的水平，以诊断由此介导的疾病状态。 此外，提供了用于诊断由这种抗性CD8T细胞亚群在慢性排斥或类风湿性关节炎中介导的持续活跃炎症的方法，其包括测量循环CD8T细胞池中生物标志物Scin的水平。

公开(公告)号：US20130189282A1

公开(公告)日：2013-07-25

申请号：US13811806

申请日：2011-07-25

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: C12N5/0783

CPC classification number: C12N5/0636 ,  C12N5/0637 ,  C12N2501/04 ,  C12N2501/06 ,  G01N33/505 ,  G01N2333/70517 ,  G01N2800/44

Abstract: Utilizing a novel T cell culture system based on allogeneic epithelial antigen presenting cells (semi-professional APC), a cyclosporin-resistant CD8 T cell clone with minimal cytolytic capability was isolated. Derivation of the novel alloantigen-specific CD8 T cell clones involved previous priming with an allogeneic skin graft, implying expansion of this T cell subset during transplant rejection. Characterization and comparison of the cyclosporin and rapamycin-resistant CD 8 T cell clone with typical cyclosporin-sensitive CD 8 T cells suggests that it is a member of a CD8 T cell subset with a unique cell surface phenotype and novel TCR activation pathways, and that these unique CD8 T cell clones reflect the immunobiology of chronic rejection within the non-hematopoetic microenvironments of solid organs and vascular walls. These cells express the aryl-hydrocarbon receptor. T-cells of this type are referred to herein as CD8bm12-1 T-cells.

Abstract translation: 利用基于同种异体上皮抗原呈递细胞（半专业APC）的新型T细胞培养系统，分离出具有最小细胞溶解能力的环孢菌素抗性CD8T细胞克隆。 衍生的新型同种异体抗原特异性CD8 T细胞克隆涉及以前用同种异体皮肤移植物引发，这意味着移植排斥期间该T细胞亚群的扩增。 环孢菌素和雷帕霉素抗性CD 8 T细胞克隆与典型的环孢菌素敏感型CD 8 T细胞的表征和比较表明它是具有独特细胞表面表型和新型TCR活化途径的CD8 T细胞亚群的成员，并且 这些独特的CD8 T细胞克隆反映了在固体器官和血管壁的非造血微环境中的慢性排斥反应的免疫生物学。 这些细胞表达芳基 - 烃受体。 这种类型的T细胞在本文中称为CD8bm12-1T细胞。

公开(公告)号：US20190302113A1

公开(公告)日：2019-10-03

申请号：US16375283

申请日：2019-04-04

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: G01N33/564 ,  G01N33/68 ,  C07K16/24 ,  C07K16/28

Abstract: Two patients diagnosed with KLS were treated. One patient had severe KLS that progressed to the equivalent of pediatric Kawasaki Disease Shock Syndrome (KDSS). The second patient had a typical KLS presentation and clinical course. Cytokines and chemokines provide inflammatory signatures in the serum that reflect the polarity of the immune response and the affected cell types. Multiplex ELISA technology was used to define the cytokine milieu in the serum of the two adult HIV patients with KLS during the acute and convalescent phases. Those sera were compared with sera from asymptomatic HIV subjects and a normal serum control. Those comparisons suggest that HIV KLS is a dysfunctional Th2 response to an unknown inciting agent in the vascular wall, and that a multiplex ELISA or similar technology based a limited combination of KLS/KD pathogenesis-related cytokines (IL-6, IL-13, sTNFRII) and endothelial/smooth muscle chemokines (CCL1, CCL2, CxCL11) may provide an objective tool for diagnosing KLS and Kawasaki Disease. Because KD and HIV KLS are the only known “Th2” vasculitidies that spare the lungs (unique clinical presentation) and include plasma cell infiltration of the vascular wall as a prominent histopathologic feature (unique pathophysiology), a diagnostic test based on combinations of the above analytes will be highly specific and therefore clinically useful.

公开(公告)号：US09185889B2

公开(公告)日：2015-11-17

申请号：US13579136

申请日：2011-02-15

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: C12N5/00 ,  A61K48/00 ,  A01K67/027 ,  C12Q1/68 ,  G01N33/15 ,  G01N33/53 ,  G01N33/68

CPC classification number: A01K67/027 ,  A01K67/0271 ,  A01K67/0276 ,  A01K2207/12 ,  A01K2217/075 ,  A01K2217/206 ,  A01K2227/105 ,  A01K2267/0337 ,  A01K2267/0387 ,  C12Q1/68 ,  G01N33/15 ,  G01N33/53 ,  G01N33/68

Abstract: MHC class II-restricted Chlamydia-specific CD4 T cell clones recognize infected upper reproductive tract epithelial cells as early as 12 hours post infection. The timing and degree of T cell activation are dependent on the interferon milieu. Different interferons have different effects on T-cell activation; interferon IFN-β blunts IFN-γ induced up regulation of epithelial cell surface MHC class II and T cell activation. A subset of CD4 T-cells that was especially good at clearing Chlamydia infections from the genital tracts of infected mice was found to express the genes Casd1 and Plac8. The mouse Casd1 genes shares some 95 percent identity with the human gene. The differential expression of either Plac8 or Casd1 in COD cells in response to an infection of epithelial tissue provides a ready methodology for the identification of individuals exposed to epithelial infections and a tool for developing vaccines against pathogens that infect epithelial tissue.

Abstract translation: MHC II类受限制的衣原体特异性CD4T细胞克隆早在感染后12小时就识别感染的上生殖道上皮细胞。 T细胞活化的时间和程度取决于干扰素环境。 不同的干扰素对T细胞活化有不同的影响; 干扰素IFN- 钝化IFN-γ诱导上皮细胞表面MHC II类和T细胞活化的调节。 发现特别擅长清除感染小鼠生殖道的衣原体感染的CD4 T细胞亚群表达基因Casd1和Plac8。 小鼠Casd1基因与人类基因共有95％的同一性。 Plac8或Casd1在COD细胞中对上皮组织感染的差异表达提供了一种用于鉴定暴露于上皮细胞感染的个体的现成方法，也是用于开发针对感染上皮组织的病原体的疫苗的工具。

公开(公告)号：US20140235543A1

公开(公告)日：2014-08-21

申请号：US14181327

申请日：2014-02-14

Applicant: Raymond M. Johnson

Inventor： Raymond M. Johnson

IPC: G01N33/569

CPC classification number: G01N33/6893 ,  G01N33/5047 ,  G01N2800/102 ,  G01N2800/24

Abstract: Methods for identifying a subset of CD8 T cells that is resistant to an inhibitor, specifically cyclosporine, rapamycin and/or tacrolimus, by detecting expression levels of human biomarkers are disclosed. The methods include determining whether a subset of certain CD8 T cells expresses elevated levels of Scin and/or Pla2g4a, with an elevated level indicative of proliferation of the identified CD8 T cells. Also disclosed are methods of diagnosing, monitoring and treating rheumatoid arthritis and transplant rejection, including determining and/or monitoring the expansion of a subset of CD8 T cells by measuring the level of expression of a biomarker in the population of the CD8 T cell subset.

Abstract translation: 公开了通过检测人类生物标志物的表达水平鉴定对抑制剂，特别是环孢菌素，雷帕霉素和/或他克莫司具有抗性的CD8T细胞亚群的方法。 所述方法包括确定特定CD8T细胞的子集是否表达升高的Scin和/或Pla2g4a水平，其中升高的水平指示鉴定的CD8T细胞的增殖。 还公开了诊断，监测和治疗类风湿性关节炎和移植排斥的方法，包括通过测量CD8T细胞亚群群体中生物标志物的表达水平来确定和/或监测CD8T细胞亚群的扩增。