Phytohemagglutinin
(Synonyms: 植物血凝素; PHA-M) 目录号 : GC36904
Phytohemagglutinin (PHA, Phaseolus vulgaris agglutinin) is expressed in Pichia pastoris using native signal peptides, or the Saccharomyces alpha-factor preprosequence, to direct proteins into the secretory pathway. Phytohemagglutinin induces apoptosis in human HEp-2 carcinoma cells via increasing proapoptotic protein Bax and activating caspases-3.
Cas No.:9008-97-3
Sample solution is provided at 25 µL, 10mM.
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Phytohemagglutinin (PHA, Phaseolus vulgaris agglutinin) is expressed in Pichia pastoris using native signal peptides, or the Saccharomyces alpha-factor preprosequence, to direct proteins into the secretory pathway. Phytohemagglutinin induces apoptosis in human HEp-2 carcinoma cells via increasing proapoptotic protein Bax and activating caspases-3.
Stimulation of human mononuclear leukocytes (MNL) by phytohemagglutinin induces the expression of ChAT mRNA, and potentiated ACh synthesis. Phytohemagglutinin binds to the membranes of T-cells, stimulates metabolic activity, cell division, and involves inflammatory pathways. [3]
Phytohemagglutinin, a standard T cell mitogen, can inhibit both T- and B-cell tumors. Phytohemagglutinin (100 μg; i.p.; daily for 2 days) inhibits A20 tumor growth in CB17-SCID mice.[4]
[1] R J Raemaekers, et al. Eur J Biochem. 1999 Oct 1;265(1):394-403. [2] T O Kochubei, et al. Exp Oncol. 2015 Jun;37(2):116-9. [3] Fujii T, et al. J Neuroimmunol. 1998 Feb;82(1):101-107.
| Cas No. | 9008-97-3 | SDF | |
| 别名 | 植物血凝素; PHA-M | ||
| Canonical SMILES | [Phytohemagglutinin] | ||
| 分子式 | 分子量 | ||
| 溶解度 | Water : 50 mg/mL | 储存条件 | 4°C, protect from light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
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2.
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Liposomal Phytohemagglutinin: In vivo T-cell activator as a novel pan-cancer immunotherapy
J Cell Mol Med 2022 Feb;26(3):940-944.PMID:35014164DOI:10.1111/jcmm.16885.
Immunotherapy is an attractive approach for treating cancer. T-cell engagers (TCEs) are a type of immunotherapy that are highly efficacious; however, they are challenged by weak T-cell activation and short persistence. Therefore, alternative solutions to induce greater activation and persistence of T cells during TCE immunotherapy is needed. Methods to activate T cells include the use of lectins, such as Phytohemagglutinin (PHA). PHA has not been used to activate T cells in vivo, for immunotherapy, due to its biological instability and toxicity. An approach to overcome the limitations of PHA while also preserving its function is needed. In this study, we report a liposomal PHA which increased PHA stability, reduced toxicity and performed as an immunotherapeutic that is able to activate T cells for the use in future cancer immunotherapies to circumvent current obstacles in immunosuppression and T-cell exhaustion.
Phytohemagglutinin isolectin subunit composition
Biochim Biophys Acta 1981 Mar 27;668(1):132-40.PMID:7236703DOI:10.1016/0005-2795(81)90156-2.
The subunit compositions of individual Phytohemagglutinin isolectins from red kidney bean Phaseolus vulgaris were examined by isoelectric focusing and sodium dodecyl sulfate electrophoresis on polyacrylamide gels. Isoelectric focusing reveals heterogeneous but unique and non-overlapping protein band patterns for each of the homotetrameric isolectins, E4 and L4. Isoelectric focusing of the intermediate isolectins which contain both subunits (E3L1, E2L2, and E1L3) show all the protein bands common to isolectins E4 or L4 in proportions relative to their suggested subunit compositions. Polyacrylamide gel electrophoresis in a continuous sodium dodecyl sulfate buffer system gives a single protein band for all of the isolectins. In contrast, a discontinuous sodium dodecyl sulfate buffer procedure resolves isolectins E4 and L4 into single major protein bands of apparent molecular weights 31 700 (+/-600) and 29 900 (+/-200), respectively. Each of the intermediate isolectins contained both protein bands and their relative proportion, as determined by absorbance scanning, confirms the Phytohemagglutinin isolectin subunit compositions as E4, E3L1, E2L2, E1L3, and L4.
T-cell response to Phytohemagglutinin in the interferon-γ release assay as a potential biomarker for the response to immune checkpoint inhibitors in patients with non-small cell lung cancer
Thorac Cancer 2021 Jun;12(11):1726-1734.PMID:33943031DOI:10.1111/1759-7714.13978.
Background: Immune checkpoint inhibitors are a standard treatment for advanced lung cancer, although it remains important to identify biomarkers that can accurately predict treatment response. Immune checkpoint inhibitors enhance the antitumor T-cell response, and interferon-γ plays an important role in this process. Therefore, this study evaluated whether the number of interferon-γ-releasing peripheral T cells after Phytohemagglutinin stimulation in the interferon-γ release assay might act as a biomarker for the response of non-small cell lung cancer to immune checkpoint inhibitor treatment. Methods: Data were retrospectively collected regarding 74 patients with non-small cell lung cancer who had received immune checkpoint inhibitors. Pretreatment screening tests had been performed using the T-SPOT.TB assay, which quantifies the number of interferon-γ-releasing T cells (as immunospots) in response to Phytohemagglutinin and tuberculosis-specific antigen stimulation. Clinical factors and the number of spots in the T-SPOT fields were evaluated for associations with patient outcomes. The median number of spots was used to categorize patients as having high or low values, and the two groups were compared. Results: Relative to patients with a low ratio, patients with a high ratio of Phytohemagglutinin/tuberculosis-specific antigen spots (i.e. more responsive T cells) had significantly better progression-free survival after immune checkpoint inhibitor treatment. When we only considered patients with negative T-SPOT results, a high number of phytohemagglutinin-stimulated spots corresponded to significantly longer progression-free survival. Conclusion: The T-SPOT.TB assay can be used to quantify the number of immunospots in response to antigen stimulation, which may predict the response to immune checkpoint inhibitors in patients with non-small cell lung cancer.
Intracellular inflammatory signalling cascades in human monocytic cells on challenge with Phytohemagglutinin and 2,4,6-trinitrophenol
Mol Cell Biochem 2022 Feb;477(2):395-414.PMID:34775567DOI:10.1007/s11010-021-04296-x.
Phytohemagglutinin (PHA) is a plant mitogen that can agglutinate human leukocytes and erythrocytes. PHA is mainly derived from red kidney beans and can act as an exogenous pyrogen. When entering into the blood circulation, exogenous pyrogens principally interact with monocytes and macrophages and induce the release of pro-inflammatory cytokines. Monocytes and macrophages are the cells that fight against foreign invaders and acts as a primary line of immune defence. Similar to PHA, the chemical 2,4,6-trinitrophenol (TNP) also acts as an exogenous pyrogen. The study focused on the in vitro interaction of PHA and TNP with the human monocyte/macrophage cell model THP-1. The exposure and associated change in cellular morphology, organelle function, mechanism of cell death, inflammatory signalling and expression of inflammation-related genes were analyzed in different time periods. It was observed that PHA and TNP induce dose and time-dependent toxicity to monocytes/macrophages where the mechanism of cell death was different for PHA and TNP. Both PHA and TNP can evoke immune signalling with increased expression of inflammatory genes and associated activation of intracellular signalling cascades.
Lymphocyte Transformation Test Based on Lymphocyte Changes Observed by a Hematology Analyzer before and after Phytohemagglutinin Stimulation
Dis Markers 2022 Nov 7;2022:5967429.PMID:36393975DOI:10.1155/2022/5967429.
Objective: The lymphocyte transformation test is a classical test for the detection of cellular immune function and is based on subjective judgment. In this study, we have established an objective novel lymphocyte transformation test using the hematology analyzer to observe lymphocyte transformation. Methods: Whole blood cells were cultured using a whole blood method with a lymphocyte culture medium; Phytohemagglutinin was used to stimulate the experimental samples, and control was set up at the same time. After the whole blood cells were cultured, the number of lymphocytes in the two groups was observed using a hematology analyzer, and the conversion rate was calculated. The new method was used to observe differences in lymphocyte conversion in the peripheral blood of patients with hematopathy and healthy persons. Results: There were significant differences between the stimulated peripheral blood group and the blank group. The transformation rate of peripheral blood lymphocytes in patients with hematopathy was significantly lower than that in healthy persons; the difference was statistically significant (P < 0.05). Conclusion: Lymphocyte transformation can be observed using a hematology analyzer. The lymphocyte transformation test that is based on the determination of lymphocyte count by a hematology analyzer has important clinical value.