ABSTRACT
The third-generation epidermal growth factor receptor (EGFR) inhibitor osimertinib (OSI) has been approved as the first-line treatment for EGFR-mutant non-small cell lung cancer (NSCLC). This study aims to explore a rational combination strategy for enhancing the OSI efficacy. In this study, OSI induced higher CD47 expression, an important anti-phagocytic immune checkpoint, via the NF-κB pathway in EGFR-mutant NSCLC HCC827 and NCI-H1975 cells. The combination treatment of OSI and the anti-CD47 antibody exhibited dramatically increasing phagocytosis in HCC827 and NCI-H1975 cells, which highly relied on the antibody-dependent cellular phagocytosis effect. Consistently, the enhanced phagocytosis index from combination treatment was reversed in CD47 knockout HCC827 cells. Meanwhile, combining the anti-CD47 antibody significantly augmented the anticancer effect of OSI in HCC827 xenograft mice model. Notably, OSI induced the surface exposure of "eat me" signal calreticulin and reduced the expression of immune-inhibitory receptor PD-L1 in cancer cells, which might contribute to the increased phagocytosis on cancer cells pretreated with OSI. In summary, these findings suggest the multidimensional regulation by OSI and encourage the further exploration of combining anti-CD47 antibody with OSI as a new strategy to enhance the anticancer efficacy in EGFR-mutant NSCLC with CD47 activation induced by OSI.
Subject(s)
Humans , Mice , Animals , Carcinoma, Non-Small-Cell Lung/metabolism , Lung Neoplasms/metabolism , Acrylamides/pharmacology , ErbB Receptors/metabolism , Cell Line, Tumor , CD47 Antigen/therapeutic useABSTRACT
【Objective】 To discuss the interference of anti-CD47 in pre-transfusion test and the mitigation measures. 【Methods】 Blood sample of one patient received anti-CD47 treatment was collected to conduct routine serological tests including ABO/Rh phenotype, direct anti-human globulin test, irregular antibody screening, antibody identification and cross-match. Packed platelet from multiple type O blood donors was used to absorb with patient′s plasma. The patient′s plasma was absorbed with CCDee, ccDEE and ccdee red cells, respectively. Anti-IgG monoclonal Gamma-clone which lacks reactivity with human subclass IgG4 was used to perform antibody screening and cross-match. Capture-R was used to perform antibody screening. 【Results】 The direct anti-human globulin test was positive(1+ ), the reactivity in all phases was strong positive(3+ -4+ ). The anti-CD47 was eliminated after platelet and red cells absorption. Antibody screening became negative using Gamma-clone and Capture-R, and cross-match successfully using Gamma-clone. 【Conclusion】 Anti-CD47 monoclonal antibody can interfere with pre-transfusion test and cross matching. To remove the interference of anti-CD47 requires the use of Gamma-clone anti-IgG in the indirect antiglobulin testing or Capture-R.
ABSTRACT
【Objective】 To discuss the case reports concerning anti-CD47 monoclonal antibody interfere in pre-transfusion testing, so as to find mitigation strategies for this drug interference. 【Methods】 Blood transfusion cases in clinical trials concerning CD47 mAb drugs at home and abroad were retrieved from PubMed, Medline, Web of Science, Wanfang data knowledge service platform and CNKI database. The characteristics and solutions of this drug interfering with pre-transfusion testing were analyzed. 【Results】 A total of 26 cases concerning anti-CD47 mAb interference in pretransfusion testing were retrieved, and 16 valid cases were included in this study (All received HU5F9-G4 as anti-CD47 mAb). After treatment with Hu5F9-G4, the discrepancy between forward and reverse blood typing reached 77% in pre-transfusion testing. Panagglutination was presented in antibody screenings, and all(100%) platelet antibody screenings was interfered. These results indicated that Hu5F9-G4 seriously affected the compatibility test of blood transfusion. Methods of eliminating anti-CD47 interference, as well as their advantages and disadvantages were further analyzed. 【Conclusion】 The advantages and disadvantages of eliminating anti-CD47 interference with pre-transfusion testing was analyzed according to its characteristics, which could provide reference for the laboratory testing.
ABSTRACT
【Objective】 To explore the impact of monoclonal anti-CD47(IBI188) on clinical pre-transfusion testing and its solutions, then compare it with monoclonal anti-CD38, so as to develop safe and rational transfusion strategies. 【Methods】 The blood typing, direct antiglobulin testing(DAT) and antibody screening were conducted by standard methods. Red blood cells(RBCs) were treated with fig protease, papain, trypsin and dithiothreitol(DTT) to observe whether the effect of monoclonal anti-CD47 could be eliminated. Cord RBCs and RBCs with different Rh phenotypes were cross-matched; Plasma samples were adsorbed with papain-treated O allogeneic RBCs. 【Results】 ABO reverse typing were affected by monoclonal anti-CD47 treatment, and all serum antibody screening were positive, and their DAT were negative or weakly positive. Neither enzyme nor DTT could weaken the effect of monoclonal anti-CD47 on antibody screening. In saline cross-matching, differences in agglutination intensity were corresponded to differences in CD47 expression on RBCs, but all RBCs agglutinated 2+ to 4+ by polybrene method and anti-human globulin method. Papain treated allogeneic RBCs can remove the monoclonal anti-CD47 in the serum through 3 to 4 rounds of absorption. 【Conclusion】 Monoclonal anti-CD47 interferes with pre-transfusion testing, which can be removed by allogeneic RBCs absorption(not suitable for antibody screening or cross-matching), but not by enzyme or DTT. Blood typing and antibody screening should be conducted before monoclonal anti-CD47 treatment and patients should be transfused with homozygous matched RBCs.