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1.
Monoclon Antib Immunodiagn Immunother ; 39(2): 45-50, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32271687

RESUMO

CD19 is a type I transmembrane glycoprotein belonging to the immunoglobulin superfamily. It is expressed in normal and neoplastic B cells, and it modulates the threshold of B cell activation for amplifying B cell receptor signaling. Blinatumomab (a CD3-CD19-bispecific T cell-engaging antibody) and tisagenlecleucel (genetically modified T cells that express a CD19 chimeric antigen receptor [CART-19]) provide significant benefits for patients with CD19-positive relapsed or refractory B cell malignancies. In this study, we first employed the Cell-Based Immunization and Screening (CBIS) method to produce anti-CD19 monoclonal antibodies using CD19-overexpressing cells for both immunization and screening. One established clone-C19Mab-1-proved to be useful in flow cytometry assays against lymphoma cell lines, such as BALL-1, P30/OHK, and Raji. Second, the extracellular domain of CD19 was immunized into mice, and enzyme-linked immunosorbent assays were performed for the first screening. One established clone-C19Mab-3-was determined to be useful for Western blotting and immunohistochemical analysis. Due to their complementary utility, a combination of C19Mab-1 (established using CBIS) and C19Mab-3 (established using conventional method) could be useful for the pathological analysis of CD19.

2.
Nat Commun ; 11(1): 1557, 2020 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-32214089

RESUMO

The telomerase reverse transcriptase is upregulated in the majority of human cancers and contributes directly to cell transformation. Here we report that hTERT is phosphorylated at threonine 249 during mitosis by the serine/threonine kinase CDK1. Clinicopathological analyses reveal that phosphorylation of hTERT at threonine 249 occurs more frequently in aggressive cancers. Using CRISPR/Cas9 genome editing, we introduce substitution mutations at threonine 249 in the endogenous hTERT locus and find that phosphorylation of threonine 249 is necessary for hTERT-mediated RNA dependent RNA polymerase (RdRP) activity but dispensable for reverse transcriptase and terminal transferase activities. Cap Analysis of Gene Expression (CAGE) demonstrates that hTERT phosphorylation at 249 regulates the expression of specific genes that are necessary for cancer cell proliferation and tumor formation. These observations indicate that phosphorylation at threonine 249 regulates hTERT RdRP and contributes to cancer progression in a telomere independent manner.


Assuntos
Proteína Quinase CDC2/metabolismo , Neoplasias/metabolismo , Neoplasias/patologia , Telomerase/metabolismo , Animais , Proteína Quinase CDC2/antagonistas & inibidores , Proteína Quinase CDC2/genética , Linhagem Celular Tumoral , Proliferação de Células , Transformação Celular Neoplásica , Regulação Neoplásica da Expressão Gênica , Células HeLa , Humanos , Camundongos , Mitose , Mutação , Neoplasias/genética , Fosforilação , RNA Replicase/metabolismo , Telomerase/genética , Treonina
3.
Spine (Phila Pa 1976) ; 45(13): E792-E798, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32044809

RESUMO

STUDY DESIGN: Case-control study. OBJECTIVE: We aimed to identify predictors for latent myelopathy and to develop a diagnostic protocol based on these factors. SUMMARY OF BACKGROUND DATA: There is no diagnostic protocol for latent myelopathy to avoid misdiagnosis in patients complaining only of lower extremity symptoms. METHODS: This case-control study identified 791 patients discussed at conferences from April 2006 to August 2012. Overall, 460 patients complaining only of lower extremity symptoms and who underwent spine surgery were included as participants; 54 underwent surgery involving the cervical and thoracic vertebrae and were assigned to the cervical-thoracic group (C-T group); 406 underwent lumbar surgery and were assigned to the lumbar group (L group). RESULTS: By univariate analysis, age ≥67 years, patellar tendon (PT) hyperreflexia, Achilles tendon (AT) hyperreflexia, spastic gait, and gait inability were more common in the C-T group than in the L group. By multivariate analysis, age ≥67 years (OR, 8; P = 0.001), AT hyperreflexia (OR, 20.5; P < 0.001), spastic gait (OR, 225; P < 0.001), and gait inability (OR, 64; P < 0.001) were significant predictive factors. In patients with age ≥67 years, PT hyperreflexia, and/or AT hyperreflexia, the sensitivity for myelopathy diagnosis was 98%. In patients with spastic gait or gait inability, the specificity of myelopathy diagnosis was 96%. CONCLUSIONS: We analyzed factors that predict latent myelopathy in patients complaining only of lower extremity symptoms. We believe a diagnostic protocol based on the predictors shown in this study would contribute to the accurate diagnosis of latent myelopathy. LEVEL OF EVIDENCE: 4.


Assuntos
Extremidade Inferior , Doenças da Medula Espinal/diagnóstico por imagem , Adulto , Idoso , Doenças da Medula Óssea , Estudos de Casos e Controles , Vértebras Cervicais/cirurgia , Erros de Diagnóstico , Feminino , Marcha , Transtornos Neurológicos da Marcha , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Doenças da Medula Espinal/cirurgia , Vértebras Torácicas
4.
Chem Asian J ; 15(5): 581-584, 2020 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-32012472

RESUMO

Irradiation of HX (X=CF3 SO3 or CF3 CO2 ) salts of 1-aryl-4-pyridylbutadienes 1 a-1 c in the solid-state afforded syn head-to-tail dimers in good yields among a number of possible dimers, whereas irradiation of the neutral substrates gave a complex mixture or no products. A comparison of the X-ray crystal structures of the neutral compounds and the HX salts clarified that their orientation modes are head-to-head and head-to-tail, respectively. Moreover, while the distances between the two neighboring double bonds of the neutral compounds are relatively far apart from each other, those of HX salts are close together, satisfying Schmidt's requirement. These findings suggested that cation-π interactions between the pyridinium and aromatic rings are effective for the preorientation of the HX salts of substrates, leading to photodimers in high regio- and stereoselectivities.

5.
Cancer Res ; 80(6): 1279-1292, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31919242

RESUMO

Pancreatic ductal adenocarcinoma (PDAC) is among the cancers with the poorest prognoses due to its highly malignant features. BTB and CNC homology 1 (BACH1) has been implicated in RAS-driven tumor formation. We focused on the role of BACH1 in PDAC, more than 90% of which have KRAS mutation. Knockdown of BACH1 in PDAC cell lines reduced cell migration and invasion, in part, by increasing E-cadherin expression, whereas its overexpression showed opposite effects. BACH1 directly repressed the expression of FOXA1 that is known to activate the expression of CDH1 encoding E-cadherin and to inhibit epithelial-to-mesenchymal transition. BACH1 also directly repressed the expression of genes important for epithelial cell adhesion including CLDN3 and CLDN4. In a mouse orthotopic implantation model, BACH1 was required for the high metastatic ability of AsPC-1 cells. IHC analysis of clinical specimens with a newly developed anti-BACH1 mAb revealed that high expression of BACH1 is a poor prognostic factor. These results suggest that the gene regulatory network of BACH1 and downstream genes including CDH1 contribute to the malignant features of PDAC by regulating epithelial-to-mesenchymal transition. SIGNIFICANCE: Greater understanding of the gene regulatory network involved in epithelial-to-mesenchymal transition of pancreatic cancer cells will provide novel therapeutic targets and diagnostic markers.

6.
Ticks Tick Borne Dis ; 11(2): 101332, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31734217

RESUMO

Tick saliva contains immunosuppressants which are important to obtain a blood meal and enhance the infectivity of tick-borne pathogens. In Japan, Ixodes persulcatus is a major vector for Lyme borreliosis pathogens, such as Borrelia garinii, as well as for those causing relapsing fever, such as B. miyamotoi. To date, little information is available on bioactive salivary molecules, produced by this tick. Thus, in this study, we identified two proteins, I. persulcatus derived sialostatin L1 (Ip-sL1) and sL2 (Ip-sL2), as orthologs of I. scapularis derived sL1 and sL2. cDNA clones of Ip-sL1 and Ip-sL2 shared a high identity with sequences of sL1 and sL2 isolated from the salivary glands of I. scapularis. Semi-quantitative PCR revealed that Ip-sL1 and Ip-sL2 were expressed in the salivary glands throughout the life of the tick. In addition, Ip-sL1 and Ip-sL2 were expressed even before the ticks started feeding, and their expression continued during blood feeding. Recombinant Ip-sL1 and Ip-sL2 were developed to characterize the proteins via biological and immunological analyses. These analyses revealed that both Ip-sL1 and Ip-sL2 had inhibitory effects on cathepsins L and S. Ip-sL1 and Ip-sL2 inhibited the production of IP-10, TNFα, and IL-6 by LPS-stimulated bone-marrow-derived dendritic cells (BMDCs). Additionally, Ip-sL1 significantly impaired BMDC maturation. Taken together, these results suggest that Ip-sL1 and Ip-sL2 confer immunosuppressive functions and appear to be involved in the transmission of pathogens by suppressing host immune responses, such as cytokine production and dendritic cell maturation. Therefore, further studies are warranted to investigate the immunosuppressive functions of Ip-sL1 and Ip-sL2 in detail to clarify their involvement in pathogen transmission via I. persulcatus.

7.
Monoclon Antib Immunodiagn Immunother ; 38(6): 282-284, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31721640

RESUMO

Podoplanin (PDPN)/T1alpha is utilized as a specific marker of lymphatic endothelial cells or type I alveolar cells of lung. Therefore, sensitive and specific monoclonal antibodies (mAbs) detecting PDPN are necessary for immunohistochemical analyses, especially using formalin-fixed paraffin-embedded tissues. Recently, we developed an anti-bear PDPN (bPDPN) mAb, PMab-247, which is useful for immunohistochemical analyses to detect both lymphatic endothelial cells and type I alveolar cells of lung. However, it is difficult to distinguish lymphatic endothelial cells from type I alveolar cells in the bear lung. In this study, we showed that a novel anti-bPDPN mAb, PMab-241 stained only lymphatic endothelial cells, not type I alveolar cells of the lung in immunohistochemical analyses. These findings suggest that PMab-241 could be useful for staining lymphatic endothelial cells specifically in the bear lung tissues.


Assuntos
Epitopos/imunologia , Pulmão/imunologia , Glicoproteínas de Membrana/imunologia , Ursidae/imunologia , Células Epiteliais Alveolares/imunologia , Animais , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos/imunologia , Células CHO , Cricetinae , Cricetulus , Humanos , Glicoproteínas de Membrana/isolamento & purificação , Podócitos/imunologia
8.
J Am Chem Soc ; 141(50): 19570-19574, 2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31736293

RESUMO

Peripherally substituted tetradecaazaheptacene (N14Hp) compounds, exhibiting amphiprotism-coupled emission, have been synthesized. X-ray crystallography reveals a planar acene-like chromophore, and electronic absorption and emission occur in the near-infrared biological transparency window (650-900 nm). The compounds exhibit long-wavelength emission with photoluminescence quantum yields ΦPL up to ∼0.61 at 686 nm, with the monodeprotonated state ΦPL ≈ 0.58 at 712 nm. This unprecedented highly nitrogenous chromophore illustrates the stability and utility of the pyrazinacenes for different applications based on their photophysical properties and chemical structures.

9.
Langmuir ; 35(48): 15784-15794, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31656076

RESUMO

The confined film structures and tribological properties of the dilute aqueous solution of a silicone-based amphiphilic block copolymer, bis-isobutyl poly(ethylene glycol) (PEG)-14/amodimethicone (BIPA) copolymer, between mica surfaces were investigated. The BIPA copolymer existed as positively charged water-soluble aggregates in the solution. The adsorption behavior of the BIPA copolymer aggregates on a mica surface from solution was studied using an atomic force microscope (AFM); the result showed the immediate formation of a uniform adsorbed BIPA copolymer layer, followed by the gradual deposition of BIPA aggregates on the top of the adsorbed layer. Friction measurements were carried out using the surface forces apparatus (SFA) for the confined films of BIPA copolymer solution between mica surfaces, which revealed two different sliding film structures depending on the elapsed time after surface preparation. The sliding film consisting of two adsorbed BIPA copolymer layers was obtained for a relatively short elapsed time (not longer than 3 h), which had an extremely low friction coefficient µ (of the order of 10-5). The sliding film on the following day (elapsed time of approximately 24 h) had the structure of a deposited/kinetically trapped BIPA aggregate layer confined between the opposing adsorbed layers, and the µ values were within the range from 10-4 to 10-3. Our results suggest that the different elapsed time ranges and resulting absence or presence of the intervening layer of trapped aggregates between the absorbed layers determine the tribological properties of the confined films. Molecular friction mechanisms are discussed for the two sliding structures, which give insight into using amphiphilic block copolymer aggregates for a new class of aqueous lubrication system to design extremely low friction interfaces.

10.
Kyobu Geka ; 72(11): 919-922, 2019 Oct.
Artigo em Japonês | MEDLINE | ID: mdl-31588109

RESUMO

Coronary artery spasm after coronary artery bypass grafting(CABG) is rare and often fatal. There is no established treatment for perioperative coronary spasm because of the rare and not fully understood complications. We report a 67-year-old male, who experienced perioperative spasm of native coronary arteries and the left thoracic artery graft following CABG. Several transcatheter intracoronary injections of isosorbide sulfate failed to relieve the spasm completely. After 5 days' circulatory support with intra-aortic balloon pumping, the cardiac function improved and the patient recovered. Echocardiography performed before discharge showed decreased left ventricular systolic function with apical akinesis. Postoperative coronary angiography revealed 4 of the 5 grafts were patent.


Assuntos
Vasoespasmo Coronário , Vasos Coronários , Idoso , Angiografia Coronária , Ponte de Artéria Coronária , Humanos , Masculino , Espasmo
11.
Monoclon Antib Immunodiagn Immunother ; 38(5): 230-233, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31535919

RESUMO

Podoplanin (PDPN)/T1alpha is a type I transmembrane sialoglycoprotein, which is expressed on podocytes of the kidneys and type I alveolar cells of the lungs. PDPN is also known as Aggrus, a platelet aggregation-inducing factor, which comprises three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) in the N-terminus and PLAG-like domains (PLDs) in the middle of the PDPN protein. We have previously established a mouse anti-bear PDPN (bPDPN) monoclonal antibody (mAb) clone, PMab-247 using the Cell-Based Immunization and Screening (CBIS) method. PMab-247 is very useful in flow cytometry, Western blotting, and immunohistochemical (IHC) analyses; however, the binding epitope of PMab-247 has not been elucidated. In this study, we aimed to investigate the epitope of PMab-247 using enzyme-linked immunosorbent assay and IHC analyses. The results revealed that the critical epitopes of PMab-247 are Asp76, Arg78, Glu80, and Arg82 of bPDPN. The Glu80 and Arg82 are included in PLD of bPDPN. The findings of our study can be applied to the production of more functional anti-bPDPN mAbs.


Assuntos
Anticorpos Monoclonais/imunologia , Epitopos/imunologia , Glicoproteínas de Membrana/imunologia , Ursidae , Animais , Anticorpos Monoclonais/metabolismo , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos/métodos , Epitopos/metabolismo , Imuno-Histoquímica , Rim/imunologia , Glicoproteínas de Membrana/genética , Mutação Puntual
12.
Monoclon Antib Immunodiagn Immunother ; 38(5): 224-229, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31464569

RESUMO

Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed on lymphatic endothelial cells, podocytes of the kidneys, and type I alveolar cells of the lungs. PDPN, a platelet aggregation-inducing factor, comprises three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) in the N-terminus and PLAG-like domains in the middle of the PDPN protein. We have previously reported a mouse antipig PDPN (pPDPN) monoclonal antibody (mAb) clone, PMab-213, which was developed using the Cell-Based Immunization and Screening (CBIS) method. PMab-213 is very useful in flow cytometry, Western blotting, and immunohistochemical analyses; however, the binding epitope of PMab-213, which was developed by CBIS method, remains to be elucidated. Therefore, this study aimed to investigate the epitope of PMab-213 using enzyme-linked immunosorbent assay, flow cytometry, and immunohistochemical analyses. The results revealed that the critical epitopes of PMab-213 are Pro53, Arg54, Arg56, and Tyr60 of pPDPN.


Assuntos
Anticorpos Monoclonais/imunologia , Epitopos/imunologia , Glicoproteínas de Membrana/imunologia , Animais , Células CHO , Cricetulus , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos/métodos , Epitopos/metabolismo , Citometria de Fluxo , Imuno-Histoquímica , Glicoproteínas de Membrana/genética , Podócitos/imunologia , Suínos
13.
J Immunol ; 203(5): 1313-1324, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31366713

RESUMO

Bovine leukemia virus (BLV) infection is a chronic viral infection of cattle and endemic in many countries, including Japan. Our previous study demonstrated that PGE2, a product of cyclooxygenase (COX) 2, suppresses Th1 responses in cattle and contributes to the progression of Johne disease, a chronic bacterial infection in cattle. However, little information is available on the association of PGE2 with chronic viral infection. Thus, we analyzed the changes in plasma PGE2 concentration during BLV infection and its effects on proviral load, viral gene transcription, Th1 responses, and disease progression. Both COX2 expression by PBMCs and plasma PGE2 concentration were higher in the infected cattle compared with uninfected cattle, and plasma PGE2 concentration was positively correlated with the proviral load. BLV Ag exposure also directly enhanced PGE2 production by PBMCs. Transcription of BLV genes was activated via PGE2 receptors EP2 and EP4, further suggesting that PGE2 contributes to disease progression. In contrast, inhibition of PGE2 production using a COX-2 inhibitor activated BLV-specific Th1 responses in vitro, as evidenced by enhanced T cell proliferation and Th1 cytokine production, and reduced BLV proviral load in vivo. Combined treatment with the COX-2 inhibitor meloxicam and anti-programmed death-ligand 1 Ab significantly reduced the BLV proviral load, suggesting a potential as a novel control method against BLV infection. Further studies using a larger number of animals are required to support the efficacy of this treatment for clinical application.


Assuntos
Anticorpos/farmacologia , Antígeno B7-H1/imunologia , Inibidores de Ciclo-Oxigenase 2/farmacologia , Dinoprostona/farmacologia , Leucose Enzoótica Bovina/tratamento farmacológico , Imunidade/efeitos dos fármacos , Vírus da Leucemia Bovina/efeitos dos fármacos , Animais , Antivirais/farmacologia , Bovinos , Ciclo-Oxigenase 2/metabolismo , Leucose Enzoótica Bovina/imunologia , Leucose Enzoótica Bovina/virologia , Vírus da Leucemia Bovina/imunologia , Provírus/efeitos dos fármacos , Provírus/imunologia , Carga Viral/efeitos dos fármacos , Carga Viral/imunologia
14.
Monoclon Antib Immunodiagn Immunother ; 38(5): 213-219, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31403389

RESUMO

Podoplanin (PDPN)/T1alpha is expressed on lymphatic endothelial cells, type I alveolar cells of the lungs, and podocytes of the kidney. PDPN possesses three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) of the N-terminus and the PLAG-like domains (PLDs). We previously reported an anti-goat PDPN (gPDPN) monoclonal antibody (mAb), PMab-235, which was developed using the Cell-Based Immunization and Screening (CBIS) method. PMab-235 is very useful in flow cytometry, Western blotting, and immunohistochemical analyses; however, the binding epitope of PMab-235 remains to be elucidated. In this study, we investigated the epitopes of PMab-235 using enzyme-linked immunosorbent assay and immunohistochemistry. The results revealed that the critical epitope of PMab-235 produced by CBIS method is Arg75, Leu78, and Pro79 of gPDPN, which is included in PLD. The findings of our study can be applied to the production of more functional anti-gPDPN mAbs.


Assuntos
Anticorpos Monoclonais/imunologia , Epitopos/imunologia , Glicoproteínas de Membrana/imunologia , Animais , Anticorpos Monoclonais/metabolismo , Colo/imunologia , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos , Epitopos/genética , Epitopos/metabolismo , Cabras , Imuno-Histoquímica , Pulmão/imunologia
15.
Monoclon Antib Immunodiagn Immunother ; 38(4): 179-182, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31403910

RESUMO

Diacylglycerol kinase (DGK) is an enzyme that converts diacylglycerol (DG) to phosphatidic acid (PA). As both DG and PA serve as lipidic second messengers, DGK plays a pivotal role in controlling the balance of two signaling pathways mediated by DG and PA in cellular functions. DGKζ, one member of the mammalian DGK family, is reported to contain a nuclear localization signal, which suggests its functional role in the nucleus. Previously, morphological studies using tagged expression vectors and immunostaining of rat tissues or cells have revealed that DGKζ localizes mainly to the nucleus. However, a limited number of studies reported the detailed localization of native protein of DGKζ in human tissues and cells. In this study, we developed a novel anti-human DGKζ monoclonal antibody, DzMab-1, which is very advantageous in immunocytochemistry of human cultured cells.


Assuntos
Anticorpos Monoclonais/imunologia , Diacilglicerol Quinase/imunologia , Hibridomas/imunologia , Imuno-Histoquímica/métodos , Animais , Células HeLa , Humanos , Imunização , Ratos , Ratos Endogâmicos WKY
16.
Heliyon ; 5(7): e02063, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31338471

RESUMO

Sensitive and specific monoclonal antibodies (mAbs) against not only human but also mouse, rat, rabbit, dog, cat, bovine, pig, and horse podoplanins (PDPNs) have been established in our previous studies. However, anti-goat PDPN (gPDPN) has not been established yet. PDPN has been utilized as a lymphatic endothelial cell marker especially in pathological diagnoses; therefore, mAbs for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues are needed. Although we recently demonstrated that an anti-bovine PDPN mAb, PMab-44 cross-reacted with gPDPN, PMab-44 did not detect lymphatic endothelial cells in immunohistochemistry. In this study, we immunized mice with gPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/gPDPN) cells, and screened mAbs against gPDPN using flow cytometry. One of the mAbs, PMab-235 (IgG1, kappa), specifically detected CHO/gPDPN cells by flow cytometry. Furthermore, PMab-235 strongly detected lung type I alveolar cells, renal podocytes, and lymphatic endothelial cells of colon by immunohistochemistry. These findings suggest that PMab-235 may be useful as a lymphatic endothelial cell marker for goat tissues.

17.
Monoclon Antib Immunodiagn Immunother ; 38(4): 171-174, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31313968

RESUMO

Podoplanin (PDPN) is utilized as a specific marker of type I alveolar cells of lung and lymphatic endothelial cells of every tissue. Therefore, sensitive and specific monoclonal antibodies detecting PDPN are necessary for immunohistochemical analyses, especially using formalin-fixed paraffin-embedded tissues. Recently, we developed an anti-bear PDPN (bPDPN) mAb, PMab-247, which is useful for Western blot, flow cytometry, and immunohistochemical analyses. In this study, immunohistochemical analyses showed that PMab-247 strongly detected bPDPN, which is expressed in type I alveolar cells and lymphatic endothelial cells of bear lung and podocytes of bear kidney. These findings suggest that PMab-247 could be useful for pathophysiological analyses using immunohistochemistry.


Assuntos
Células Epiteliais Alveolares/metabolismo , Anticorpos Monoclonais/imunologia , Células Endoteliais/metabolismo , Rim/metabolismo , Pulmão/metabolismo , Glicoproteínas de Membrana/imunologia , Células Epiteliais Alveolares/imunologia , Animais , Células Endoteliais/imunologia , Imuno-Histoquímica/métodos , Rim/imunologia , Pulmão/imunologia , Ursidae
18.
Monoclon Antib Immunodiagn Immunother ; 38(3): 104-107, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31161964

RESUMO

Podoplanin (PDPN)/T1 alpha is known as a specific marker of lymphatic endothelial cells and type I alveolar cells. Sensitive and specific monoclonal antibodies (mAbs) for PDPN are needed for immunohistochemical analyses. Recently, we developed an anticetacean PDPN mAb, PMab-237. Herein, immunohistochemical analyses showed that PMab-237 strongly detected pulmonary type I alveolar cells, renal podocytes, and lymphatic endothelial cells of the harbor porpoise. These findings suggest that PMab-237 may be useful for immunohistochemical analyses for cetacean tissues.


Assuntos
Células Epiteliais Alveolares/metabolismo , Anticorpos Monoclonais/imunologia , Células Endoteliais/metabolismo , Epitopos/imunologia , Glicoproteínas de Membrana/imunologia , Phocoena/metabolismo , Podócitos/metabolismo , Células Epiteliais Alveolares/imunologia , Animais , Especificidade de Anticorpos/imunologia , Células Endoteliais/imunologia , Humanos , Imuno-Histoquímica , Phocoena/imunologia , Podócitos/imunologia
19.
Monoclon Antib Immunodiagn Immunother ; 38(3): 108-113, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31161965

RESUMO

Podoplanin (PDPN) has been utilized as a lymphatic endothelial cell marker especially in pathological diagnoses. Therefore, sensitive and specific monoclonal antibodies (mAbs) targeting PDPN are needed for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues. Recently, anti-PDPN mAbs against many species, such as human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse were established in our studies. However, anticetacean (whale) PDPN (wPDPN) has not been established yet. In this study, we immunized mice with wPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/wPDPN) cells, and screened mAbs against wPDPN using flow cytometry. One of the mAbs, PMab-237 (IgG1, kappa), specifically detected CHO/wPDPN cells by flow cytometry and immunohistochemistry. Our findings suggest the potential usefulness of PMab-237 for the functional analyses of wPDPN.


Assuntos
Anticorpos Monoclonais/imunologia , Epitopos/imunologia , Glicoproteínas de Membrana/imunologia , Phocoena/metabolismo , Animais , Especificidade de Anticorpos , Células CHO , Gatos , Bovinos , Cricetinae , Cricetulus , Feminino , Cavalos , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos BALB C , Phocoena/imunologia , Coelhos , Ratos , Suínos
20.
Monoclon Antib Immunodiagn Immunother ; 38(4): 157-161, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31199696

RESUMO

The expression of human epidermal growth factor receptor 2 (HER2) has been reported to be overexpressed in several cancers, such as breast, lung, gastric, pancreatic, and colorectal cancers, and be associated with poor clinical outcomes. Trastuzumab, a humanized anti-HER2 antibody, provides significant survival benefits for patients with HER2-overexpressing breast cancers and gastric cancers. In this study, we developed a novel anti-HER2 monoclonal antibody (mAb), H2Mab-41 (IgG2b, kappa), and the antitumor activity of H2Mab-41 was investigated using mouse xenograft models. Caco-2 cells (human colon cancer cell line), which expresses HER2, were subcutaneously implanted into the flanks of nude mice. H2Mab-41 and control mouse IgG were injected three times into the peritoneal cavity of mice. H2Mab-41 significantly reduced tumor development of Caco-2 xenograft in comparison with the control mouse IgG on days 5, 8, 11, 15, and 19. Taken together, these results suggest that H2Mab-41 is useful for antibody therapy against HER2-expressing colon cancers.


Assuntos
Anticorpos Monoclonais/farmacologia , Neoplasias do Colo/prevenção & controle , Receptor ErbB-2/antagonistas & inibidores , Receptor ErbB-2/imunologia , Animais , Anticorpos Monoclonais/imunologia , Apoptose , Proliferação de Células , Neoplasias do Colo/imunologia , Neoplasias do Colo/patologia , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
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