Cell Lymphocyte Proliferation Research Articles

Magnetic resonance imaging-based approaches for detecting the efficacy of combining therapy following VEGFR-2 and PD-1 blockade in a colon cancer model

BackgroundAngiogenesis inhibitors have been identified to improve the efficacy of immunotherapy in recent studies. However, the delayed therapeutic effect of immunotherapy poses challenges in treatment planning. Therefore, this study aims to explore the potential of non-invasive imaging techniques, specifically intravoxel-incoherent-motion diffusion-weighted imaging (IVIM-DWI) and blood oxygenation level-dependent magnetic resonance imaging (BOLD-MRI), in detecting the anti-tumor response to the combination therapy involving immune checkpoint blockade therapy and anti-angiogenesis therapy in a tumor-bearing animal model.MethodsThe C57BL/6 mice were implanted with murine MC-38 cells to establish colon cancer xenograft model, and randomly divided into the control group, anti-PD-1 therapy group, and combination therapy group (VEGFR-2 inhibitor combined with anti-PD-1 antibody treatment). All mice were imaged before and, on the 3rd, 6th, 9th, and 12th day after administration, and pathological examinations were conducted at the same time points.ResultsThe combination therapy group effectively suppressed tumor growth, exhibiting a significantly higher tumor inhibition rate of 69.96% compared to the anti-PD-1 group (56.71%). The f value and D* value of IVIM-DWI exhibit advantages in reflecting tumor angiogenesis. The D* value showed the highest correlation with CD31 (r = 0.702, P = 0.001), and the f value demonstrated the closest correlation with vessel maturity (r = 0.693, P = 0.001). While the BOLD-MRI parameter, R2* value, shows the highest correlation with Hif-1α(r = 0.778, P < 0.001), indicating the capability of BOLD-MRI to evaluate tumor hypoxia. In addition, the D value of IVIM-DWI is closely related to tumor cell proliferation, apoptosis, and infiltration of lymphocytes. The D value was highly correlated with Ki-67 (r = − 0.792, P < 0.001), TUNEL (r = 0.910, P < 0.001) and CD8a (r = 0.918, P < 0.001).ConclusionsThe combination of VEGFR-2 inhibitors with PD-1 immunotherapy shows a synergistic anti-tumor effect on the mouse colon cancer model. IVIM-DWI and BOLD-MRI are expected to be used as non-invasive approaches to provide imaging-based evidence for tumor response detection and efficacy evaluation.

Total Phenolic Content and Proliferation Activity of Spirulina Extract in Lymphocyte Cell

Decreased immunity can make the body susceptible to diseases. Many side effects due to the use of drugs such as chemotherapy drugs cause many people to use natural ingredients to be used as supplements. Lymphocyte cell proliferation is cell division or multiplication to enhance the immune system. This study aims to determine the total phenolic content of the 70% ethanolic extract of Spirulina platensis and to analyze the potential of this extract to increase the proliferative activity of lymphocyte cells. The measurement of total phenolic levels used the Folin-Ciocalteu method. The measurement of the viability of lymphocyte cell proliferation activity used the MTT method. The 70% ethanolic extract of S. platensis in this study had a total phenolic content of 2.4957±0.0597 GAE mg/g. The 70% ethanolic extract of S. platensis increased the lymphocyte cell proliferation activity. The best viability results were found at a concentration of 20 ppm of 124.89±1.84%.

Curcumin Enhances Ibrutinib Induced Killing Effect Against TP53-Mutated Chronic Lymphocytic Leukemia Cells in Vitro

Introduction TP53 mutation and del (17p) are the most important poor prognostic factors in patients with Chronic Lymphocytic Leukemia (CLL).Patients with deletion 17p or TP53 mutations tend to have lower response rates to standard first-line chemoimmunotherapy and a more aggressive clinical course(Stilgenbauer et. al. Blood 2014). RESONATE-2 study long-term follow-up showing that ibrutinib continues to demonstrate significant and durable clinical benefit in older patients, including those with TP53 mutation(Barr et. al. Blood Adv 2022).However, CLL bearing del(17p) showed inferior OS and time to next treatment compared to non-del(17p)(Mato et. al. Haematologica 2022). A significant proportion of patients treated with single-agent ibrutinib experienced CLL progression.BTK and/or PLCG2 mutations were present in 80.0% of patients progressing with CLL, confirming prior reports on the high prevalence of these mutations in patients with ibrutinib-resistant CLL(Ahn et. al. Blood 2017). Curcumin is a bioactive natural polyphenol compound extracted from the Curcuma longa(Zingiberaceae plant). Curcumin has anti-tumor effect and can induce apoptosis and inhibit the proliferation of a series of tumor cells(Willenbacher et. al. Int J Mol Sci 2019). Curcumin may limit the development of therapy-related toxicity and reduce the financial burden of treatment.In this study, we found that Curcumin inhibits TP53-mutated CLL cells, and eliciting a strong synergistic cytotoxic effect in combination with ibrutinib. Methods The inhibitory effect of ibrutinib combined with curcumin on the proliferation of TP53-mutated Chronic Lymphocytic Leukemia cells was detected by CCK8, and the apoptosis of TP53-mutated CLL cells induced by ibrutinib combined with curcumin was detected by flow cytometry;The differences of gene and signal pathway expression among different treatment groups were analyzed by RNA-seq ;Using Western blot technology to verify the expression of related pathway proteins and explore the mechanism of curcumin increasing the killing effect of ibrutinib against TP53-mutated Chronic Lymphocytic Leukemia cells. Results The CCK8 results showed that the IC50 of ibrutinib for 24 hours was 40.61±2.35μM, while the IC50 of curcumin for 24 hours was 26.6±0.94μM. The IC50 of ibrutinib for 48 h was 14.6 ±0.8307μM, and the IC50 of curcumin for 48 h was 15.7±2.53μM.Cell inhibition of various group for 48h is shown in Figure 1.The CI of ibrutinib combined with curcumin at 24 hours and 48 hours was calculated by CompuSyn software. The results showed that the CI values of ibrutinib combined with curcumin were less than 1 at 24 h and 48 h, suggesting that ibrutinib combined with curcumin has a synergistic killing effect on CLL cells with TP53 mutation;Flow cytometry showed that the IC50 of ibrutinib for 24 hours was 67.20±2.86μM, while the IC50 of curcumin for 24 hours was 144±6.6 μM. The CI value of ibrutinib combined with curcumin was less than 1 at 24 h, suggesting that ibrutinib combined with curcumin has a synergistic promoting apoptosis on CLL cells with TP53 mutation;RNA-seq analysis and KEGG analysis of down-regulated genes of combination group compared with the control group were mainly enrich in PI3K/AKT, Cytokine-Cytokine Recptor signal pathway and so on;4.The results of Western blot showed that the protein expression levels of P65PI3K and p53 in the single drug group and the combination group were down-regulated in varying degrees, especially in the combination group（Figure 2). Conclusion Ibrutinib combined with curcumin has synergistic killing effect on TP53- mutated CLL cells;Curcumin increases the killing effect of ibrutinib on TP53-mutated CLL cells by inhibiting PI3K and NFκB pathway, and degrading p53mt protein.

Immunostimulant Compounds Identification in Indonesian Underutilized Zingiberaceae Spices

Spices are known for its health beneficial effect beside its function as flavouring and condiment. This study aimed to screen immunostimulant activities of 12 underutilized species of Zingiberaceae grown in Indonesia. The compound associated with immunostimulant activity in the most active sample was identified by linking the chromatogram profile of hexane, chloroform, ethyl acetate, methanol, and water fractions of the selected spices to its activity using orthogonal projection to the least square (OPLS) analysis, followed by LC-MS characterization. The result showed that Alpinia malaccensis and Zingiber ottensii had the highest lymphocyte cell proliferation index (SI) at concentrations of 400 ppm. A. malaccensis was chosen for the next step. The results of OPLS analysis showed that the retention time interval which strongly correlated with the immunostimulant activity was 27.0 – 28.5 minutes, which were abundant in methanol fraction. The mass spectra analysis of methanol fraction using UHPLC-HRMS showed that the respective retention time was attributed to flavokawain B. This compound was reported to increase rat splenocyte proliferation in vivo.

Preferential differential gene expression within the WC1.1+ γδ T cell compartment in cattle naturally infected with Mycobacterium bovis.

Bovine tuberculosis (bTB), caused by infection with Mycobacterium bovis, continues to cause significant issues for the global agriculture industry as well as for human health. An incomplete understanding of the host immune response contributes to the challenges of control and eradication of this zoonotic disease. In this study, high-throughput bulk RNA sequencing (RNA-seq) was used to characterise differential gene expression in γδ T cells - a subgroup of T cells that bridge innate and adaptive immunity and have known anti-mycobacterial response mechanisms. γδ T cell subsets are classified based on expression of a pathogen-recognition receptor known as Workshop Cluster 1 (WC1) and we hypothesised that bTB disease may alter the phenotype and function of specific γδ T cell subsets. Peripheral blood was collected from naturally M. bovis-infected (positive for single intradermal comparative tuberculin test (SICTT) and IFN-γ ELISA) and age- and sex-matched, non-infected control Holstein-Friesian cattle. γδ T subsets were isolated using fluorescence activated cell sorting (n = 10-12 per group) and high-quality RNA extracted from each purified lymphocyte subset (WC1.1+, WC1.2+, WC1- and γδ-) was used to generate transcriptomes using bulk RNA-seq (n = 6 per group, representing a total of 48 RNA-seq libraries). Relatively low numbers of differentially expressed genes (DEGs) were observed between most cell subsets; however, 189 genes were significantly differentially expressed in the M. bovis-infected compared to the control groups for the WC1.1+ γδ T cell compartment (absolute log2 FC ≥ 1.5 and FDR P adj. ≤ 0.1). The majority of these DEGs (168) were significantly increased in expression in cells from the bTB+ cattle and included genes encoding transcription factors (TBX21 and EOMES), chemokine receptors (CCR5 and CCR7), granzymes (GZMA, GZMM, and GZMH) and multiple killer cell immunoglobulin-like receptor (KIR) proteins indicating cytotoxic functions. Biological pathway overrepresentation analysis revealed enrichment of genes with multiple immune functions including cell activation, proliferation, chemotaxis, and cytotoxicity of lymphocytes. In conclusion, γδ T cells have important inflammatory and regulatory functions in cattle, and we provide evidence for preferential differential activation of the WC1.1+ specific subset in cattle naturally infected with M. bovis.

The Extract of Pinellia Ternata-Induced Apoptosis of Leukemia Cells by Regulating the Expression of Bax, Bcl-2 and Caspase-3 Protein Expression in Mice

The study aims to lessen the monetary burden on patients and society by decreasing the price of proprietary drugs used in leukemia therapy. Flow cytometry, reverse transcription polymerase chain reaction, western blot, and a patient-derived xenograft mouse model were used to confirm the therapeutic effect of Pinellia ternata extract on leukemia. Three types of leukemia cells (K562, HL-60, and C8166 cell lines) were found to undergo early apoptosis (P ≤ .05) after being exposed to P. ternata extract, as measured by flow cytometry. Reverse transcription polymerase chain reaction results showed that P. ternata extract at both middle (300 μg/mL) and high (500 μg/mL) concentrations was able to down-regulate Bcl-2 and upregulate mRNA expression of Bax and caspase-3. In the patient-derived xenograft mouse model formed by BALB/c-nu/nu nude mice, immunohistochemistry indicated that P. ternata extract effectively suppressed the proliferation of leukemia cells. Therefore, P. ternata extract at 300 μg/mL and 500 μg/mL could effectively inhibit myeloid and lymphocytic leukemia cell proliferation and promote leukemia cell apoptosis by regulating Bax/Bcl-2 and caspase-3.

Intracellular Potassium in Cell Growth and Proliferation of Human Mesenchymal Stem Cells and Blood Lymphocytes

Many data show that K+ ions are essential for cell proliferation. In this brief review, we summarize our own studies and literature data that characterize the relationship between modulations of intracellular K+ content and the intensity of cell proliferation. Using flame emission assay we compared the transport of monovalent cations in transformed cells and in human mesenchymal stem cells in growing cultures, as well as during stress-induced cell cycle arrest and transition from monolayer (2D) to three-dimensional (3D) spheroids. A decrease in the intracellular content of K+ (evaluated as the ratio of the content of K+ in cells to the mass of cellular protein) associated with the accumulation of G1 cells in the population and accompanied by a stop in proliferation was revealed. The relationship between intracellular K+ and initiation of cell proliferation was further analyzed in human blood lymphocytes (HBL) as a model for the transition of cells from quiescence to proliferation. In HBL stimulated to proliferate, the content of K+ in cells increases during the transition G0/G1/S, accompanying the growth of small lymphocytes into blasts. Cellular water content, calculated from buoyant cell density, is higher in proliferating HBLs and in Jurkat leukemic T cells than in resting HBL. Available data suggest that intracellular K+ is important for successful cell proliferation as the main intracellular ion involved in the regulation of cell volume during the transition from quiescence to proliferation, and high K+ content and associated high water content in the cell are a characteristic feature of cell proliferation and transformation.

Immunomodulatory Effect of Ethanol Extract of Pirdot Leaf (Saurauia vulcani Korth ) on CD4+ and IL-2 Hepatic Profile in Tyfoid Fever

Typhoid fever is an acute febrile disease caused by Salmonella typhi bacteria that is obtained orally either through food or from drinks.Many immunological activities that occur due to salmonella infection so that immunomodulators are needed to speed up the healing of one of them by using pirdot leaves. Pirdot leaves are widely distributed in Indonesia, especially in the Lake Toba area of North Sumatra.This plant has many bioactive components, one of which is flavonoids.The flavonoid component of pirdot Leaf has been widely studied and is believed to improve the immune system because it is able to increase the production of IL-2 which is involved in the activation and proliferation of lymphocyte cells and can increase T CD4 cells. T CD4+ it self is a signal transduction marker in the mechanism of typhoid fever that can activate Antigen Precenting Cell (APC) to provide resistance to foreign substances.

Aktivitas Imunostimulan Ekstrak Metanol Biji Kurma (Phoenix dactylifera) secara In Silico Terhadap Reseptor GIF dan COX-2 serta Uji In Vitro melalui Proliferasi Sel Limfosit Mencit

Dates (Phoenix dactylifera) is one of the plants that very useful, but the date palm seeds that become waste have not been widely used. Based on studies that are known to contain bioactive compounds that can be suspected as antioxidants, antimicrobials and anti-inflammatory. However, research on immunostimulants from dates, especially in silico and in vitro, has not been widely carried out. Therefore, this study aims to determine the potential of date seed bioactive compounds as immunostimulants through in silico against Human Glycosylation-Inhibiting Factor (1GIF) and Human Cyclooxygenase-2 (5IKR) receptors as well as in vitro through the proliferation of lymphocyte cells of male mice (Mus musculus) balb/c strains with the MTT Assay methods. The results of the in silico study showed that 5-O-Caffeoylshikimic Acid had the highest interaction and stability, Gibbs free energy with 1GIF and 5IKR receptors, -6.68 kcal/mol and -8.58 kcal/mol, respectively. This value is known to be higher than the Caffeic Acid as control ligand. The results of in vitro research obtained methanol extract of date seeds at a concentration series of 6.25; 12.5; 25 µg/mL had higher lymphocyte cell proliferation activity which was indicated by an increasing OD value, and at a concentration of 25 μg/mL produces the highest immunostimulant activity. Therefore, date seed extract containing the compound 5-O-Caffeoylshikimic Acid can potentially be an immunostimulant agent.

Potential of Fig Leaf (Ficus carica ) as Immunomodulatory Agent in vitro and in Silico

Natural remedies are increasingly being used as alternatives to conventional medicine, and their popularity is growing. The fig plant (Ficus carica L.), which is known to contain secondary metabolites such as alkaloids, flavonoids, phenolics, terpenoids, steroids, and saponins, has the potential to act as an immunomodulator. The purpose of this study was to determine the content of flavonoid compounds based on Thin Layer Chromatography (TLC), total flavonoid content, total phenolic content, and immunomodulatory activity of ethanol extract and fig leaf fraction in vitro and silico (molecular docking). The fig leaf powder was extracted with 70% ethanol and fractionated with n-hexane and ethyl acetate to obtain ethanol extract, ethyl acetate fraction, n-hexane fraction, and water fraction of fig leaves. The extracts and fractions were then identified using the TLC method, and the total flavonoid and total phenolic levels were measured using the colorimetric method. Furthermore, the proliferation of lymphocyte cells and macrophage phagocytic activity were used to gauge the in vitro immunomodulatory activity. The results showed that the ethyl acetate fraction sample contained the highest total flavonoid and total phenolic content, namely 3.4630±0.04 mgQE/g sample and 220.1801±0.604 mgGAE/g sample. The immunomodulatory activity test findings revealed that fig leaf extract and fraction might enhance macrophage phagocytic activity in comparison to control cells. In the lymphocyte proliferation test, the value of IS>3 in the ethyl acetate fraction indicates that it has lymphocyte proliferation activity. This study showed that fig leaf extract and fraction could increase the phagocytic activity of macrophage cells. The ethyl acetate fraction of fig leaf could increase lymphocyte cell proliferation in vitro and silico.

In Vitro Effects of Low-energy Ultrasound Treatment on Healthy CD3/CD8+ Lymphocytes, Red blood cells, Acute Myeloid leukemia cells, and Jurkat cell line.

The study of the biological effects of low-energy ultrasound and its applications is a rapidly expanding research area. Low-energy ultrasound could be used as anti-tumoral therapy with or without the pharmacological combination even if the second situation has been scarcely investigated up to now. Very little information is available about the ultrasound effects on healthy red blood cells, CD3, and mainly CD8 subset lymphocytes which is the main subset cell having cytotoxic function towards cancer cells. In this study, we investigated in vitro the bioeffects of low energy ultrasound on red blood cells and PBMCs isolated from healthy donors as well as on two myeloid leukemia cell lines (OCI- AML-3 MOLM-13) and lymphoblastic Jurkat cell line. Using low-energy ultrasound (US), a study was conducted to determine how it affects CD3/CD8 lymphocytes and leukemia cells, as well as its potential role in treating blood cancers, by analyzing changes in mitochondrial membrane potential, phosphatidylserine asymmetry, morphological changes for myeloid AML cell lines, proliferation and cytotoxic activation of healthy lymphocytes, and apoptosis for RBCs after US exposure. Overall, we demonstrated that CD3/CD8 lymphocytes proliferation/activation and cytotoxic functions are fully preserved after ultrasound treatments, whereas leukemia cell lines undergo apoptosis and stop proliferating suggesting a potential method of treating blood cancer.

The eye and lymphoma: New concepts of an old acquaintance

AbstractIntraocular lymphoma (IOL) is a life‐threatening condition that results from the proliferation of malignant lymphocytic cells arising from either the central nervous system (CNS) (primary CNS lymphoma) or outside the CNS with secondary dissemination and metastasis to the eye (secondary IOL). Primary intraocular lymphoma (PIOL), also known as primary vitreoretinal lymphoma (PVRL), is a subset of PCNSL where the findings are ocular.PVRL is particularly feared because it can manifest as non‐specific uveitis, leading to a potentially fatal diagnostic delay. Although clinical and ancillary parameters can provide valuable diagnostic clues, the only valid confirmation comes from cytological analysis and molecular studies of clonality based on vitreous and/or (sub)retinal biopsies performed during vitrectomy. Standard three‐port twenty‐five‐gauge transconjunctival vitrectomy is the usual sampling technique; however, it is important to remember that the diagnostic yield of vitreous cells depends on the way samples are harvested, transported and analysed. Even when surgical recommendations are strictly followed and analyses performed by trained pathologists, the diagnostic yield of vitrectomy is not 100%.The objective of this course is to describe the clinical features of PVRL and its characteristics on ancillary exams in both typical and more challenging situations, and present the diagnostic procedures that are or will be available in the near future. Therapeutic management will be discussed later during this course.

Increased oxidative and chromosomal DNA damage in patients with ankylosing spondylitis: its role in pathogenesis.

Increased DNA damage has been suggested to contribute to the pathogenesis of chronic inflammatory diseases, but controlled studies are lacking in ankylosing spondylitis (AS). Therefore, we assessed oxidative stress, oxidative DNA damage, chromosomal DNA damage, cell proliferation and cell death in the peripheral blood lymphocytes of patients with AS as well as the possible role of DNA damage in the development of the disease. In total, 25 newly diagnosed AS patients who had not received anti-inflammatory agents and 25 healthy controls were recruited. Oxidative DNA damage was assessed by plasma 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, and chromosomal DNA damage was assessed by the cytokinesis-block micronucleus cytome (CBMN-cyt) method. Compared to controls, the micronucleus (MN) frequencies, nucleoplasmic bridge (NPB) frequencies, nuclear bud (NBUD) frequencies, apoptotic cell frequencies, necrotic cell frequencies and plasma 8-OHdG levels were significantly higher in patients with AS (p < 0.001, p < 0.05, p < 0.01, p < 0.001, p < 0.001, and p < 0.001, respectively), and the metaphase cell numbers, binucleated (BN) cell frequencies and nuclear division index (NDI) values were significantly lower in patients with AS (p < 0.01, p < 0.001 and p < 0.001, respectively). Thus, the present findings suggested that oxidative stress, oxidative DNA damage, and chromosomal DNA damage may be involved in the pathogenesis of AS similar to other chronic inflammatory diseases. In addition, the increased plasma 8-OHdG levels, MN frequencies, NPB frequencies and NBUD frequencies in AS patients may reflect an increased cancer risk.

In Vitro Immunomodulatory Activity of Fig Fruit Ethanol Extract (Ficus carica Linn) against Phagocytosis Macrophages and Lymphocyte Proliferation

Fig (Ficus carica L.) is a natural product that potentially can improve the immune system because it has flavonoids that have the potential as immunostimulants. The research aims to determine the possibility of fig fruit ethanol extract as an immunomodulator. Immunomodulatory activity is determined by knowing the activity of macrophage phagocytosis and lymphocyte proliferation in vitro and the levels of flavonoids in the extract. The research began with extraction, and then the sample was tested with TLC and colorimetry methods. Furthermore, the sample in the immunomodulatory activity test in vitro was measured through the activity of macrophage phagocytosis and lymphocyte proliferation. In the phagocytosis activity test, macrophage cells were given samples in various concentrations and latex beads. The number of activated macrophages and the number of latex phagocyted by the macrophage is then calculated. For tests of lymphocyte proliferation activity, lymphocyte cells were sampled with different concentrations and induced hepatitis B vaccine. Then the cell absorbance was read with an Elisa reader at 550nm wavelength. The study results found that the samples contained flavonoid compounds, and the total flavonoid levels obtained were 0.74±0.01 mgEQ/g samples. The immunomodulatory activity showed that the sample increased phagocytosis activity of macrophages compared to cell control. The lymphocyte proliferation test produced stimulation index&lt;2 values, showing no effect on the proliferation of lymphocytes. This study indicated that fig fruit ethanol extract could increase the phagocytosis activity of macrophage cells but did not affect the proliferation of lymphocyte cells in vitro.

​​New Approach for Development of Inactivated RVF Virus Vaccine by using Chitosan and Aluminum Phosphate Nanoparticles as Adjuvant

Background: Rift Valley fever (RVF) is a zoonotic disease that causing great economic losses in animals wealth. Vaccination with RFV vaccines are an important factor for controlling the disease. This study was conducted to investigate the immunological effect of chitosan and aluminum phosphate nanoparticles as adjuvants. Methods: Evaluating the cellular and humoral immune responses in sheep vaccinated with the prepared RVF vaccines. Result: The prepared vaccines were sterile and safe. The potent lymphocytic cell proliferation of cell-mediated immune response was early increased with significant level from 1st day post-inoculation and reached the peak at the 5th, 7th and 10th days post-inoculation in groups vaccinated with Chitosan nanoparticles RVF vaccine, aluminum phosphate nanoparticles vaccine and aluminum hydroxide RVF vaccine respectively. The results of cytokines (IL-12, TNF-α mRNA expression level and IFN-γ) were consistent with the results of cell proliferation. Neutralizing antibody it was increased significantly from the 2nd week post-inoculation and reached the peak at 5th, 4th and 3rd months in groups inoculated with Chitosan nanoparticles RVF vaccine, aluminum phosphate nanoparticles vaccine and aluminum hydroxide RVF vaccine respectively. ELISA results correlated with that obtained by SNT. Our study showed that both nanoparticles RVF vaccines induced rapid immunological response with long duration of immunity.

In Vitro Immunomodulatory Activity of Virgin Coconut Oil (VCO) with and without Bromelain Enzyme from Pineapple Waste (Ananas comosus (L) Merr)

Immunomodulators are an essential part of the prevention process for treating various diseases related to the body’s immune system. This study aimed to determine the immunomodulatory activity of virgin coconut oil (VCO) with and without the enzyme bromelain in pineapple waste extract on the proliferation of mice lymphocyte cells through an in vitro test. VCO was made using two methods: enzymatic using bromelain enzyme (VCOb) from pineapple waste with 10, 25, and 50%, and mixing method without bromelain enzyme (VCOm). The two types of VCO produced were calculated for the yield, moisture content, free fatty acids (FFA), and physicochemical properties. For immunomodulatory activity, the test solution was taken from VCOb and VCOm with a variation concentration of 6.25, 12.5, 25, 50, and 100 µg/mL. Isolation of lymphocyte cells was obtained from the spleen organ of Swiss Webster strain mice which was performed aseptically with a ketamine-xylazine anesthetic. The lymphocyte cell proliferation test was performed using the MTT Assay, and the Optical Density was measured using a microplate reader at 550 nm. The results showed that 50% VCOb produced the highest yield and 22.22% higher than VCOm. However, the results showed that increasing bromelain enzyme concentration would increase the moisture content and free fatty acid content, but still below 0.2%. The VCOb and VCOm had immunomodulatory activity against mice lymphocyte cell proliferation. However, the immunomodulatory activity of VCO with bromelain enzyme from pineapple waste extract (VCOb) was higher than without bromelain enzyme (VCOm). The highest immunomodulatory activity was obtained at 100 µg/mL of VCOb with a percentage increase of 158.26% compared to negative controls, followed by VCOm of 100 µg/mL with a percentage increase of 137.66% compared to negative controls. The optimum dose of VCOm and VCOb for increasing the proliferation of mice lymphocyte cells has not been found.

Explainable AI in Diagnosing and Anticipating Leukemia Using Transfer Learning Method

White blood cells (WBCs) are blood cells that fight infections and diseases as a part of the immune system. They are also known as “defender cells.” But the imbalance in the number of WBCs in the blood can be hazardous. Leukemia is the most common blood cancer caused by an overabundance of WBCs in the immune system. Acute lymphocytic leukemia (ALL) usually occurs when the bone marrow creates many immature WBCs that destroy healthy cells. People of all ages, including children and adolescents, can be affected by ALL. The rapid proliferation of atypical lymphocyte cells can cause a reduction in new blood cells and increase the chances of death in patients. Therefore, early and precise cancer detection can help with better therapy and a higher survival probability in the case of leukemia. However, diagnosing ALL is time-consuming and complicated, and manual analysis is expensive, with subjective and error-prone outcomes. Thus, detecting normal and malignant cells reliably and accurately is crucial. For this reason, automatic detection using computer-aided diagnostic models can help doctors effectively detect early leukemia. The entire approach may be automated using image processing techniques, reducing physicians' workload and increasing diagnosis accuracy. The impact of deep learning (DL) on medical research has recently proven quite beneficial, offering new avenues and possibilities in the healthcare domain for diagnostic techniques. However, to make that happen soon in DL, the entire community must overcome the explainability limit. Because of the black box operation's shortcomings in artificial intelligence (AI) models' decisions, there is a lack of liability and trust in the outcomes. But explainable artificial intelligence (XAI) can solve this problem by interpreting the predictions of AI systems. This study emphasizes leukemia, specifically ALL. The proposed strategy recognizes acute lymphoblastic leukemia as an automated procedure that applies different transfer learning models to classify ALL. Hence, using local interpretable model-agnostic explanations (LIME) to assure validity and reliability, this method also explains the cause of a specific classification. The proposed method achieved 98.38% accuracy with the InceptionV3 model. Experimental results were found between different transfer learning methods, including ResNet101V2, VGG19, and InceptionResNetV2, later verified with the LIME algorithm for XAI, where the proposed method performed the best. The obtained results and their reliability demonstrate that it can be preferred in identifying ALL, which will assist medical examiners.

Taraxasterol mitigates Con A-induced hepatitis in mice by suppressing interleukin-2 expression and its signaling in T lymphocytes

Discovery of anti-inflammatory drugs that can suppress T lymphocyte activation and proliferation by inhibiting TCR/CD3 and IL-2/IL-2R signaling is still needed in clinic, though rapamycin and other related reagents have made great success. Taraxasterol (TAS) is an active ingredient of dandelion, an anti-inflammatory medicinal herb with low in vivo toxicity that has long been used in China. Yet the action mechanism of TAS on lymphocytes remains elusive. The anti-inflammatory effects of TAS were evaluated in C57BL/6 mouse primary lymphocytes stimulated with concanavalin A (Con A) in vitro and in mouse model of Con A-induced acute hepatitis in vivo. Our results showed that TAS significantly suppressed Con A-induced acute hepatitis in a mouse model, reducing the hepatic necrosis areas, the release of aminotransferases, and the production of IL-2 and other inflammatory cytokines. Supporting this, in vitro study also showed that TAS reduced the production of IL-2 and the expression of IL-2 receptor subunit α (CD25) upon the stimulation of Con A, which was likely mediated by suppressing NF-κB activation. The downstream pathways of IL-2/IL-2R signaling, including the activation of PI3K/PDK1/mTOR, STAT3 and STAT5, were also suppressed by TAS. Consistently, Con A-induced T cell proliferation was also inhibited by TAS in vitro. Our data indicate that TAS can suppress both T lymphocyte activation and cell proliferation by down-regulating IL-2 expression and its signaling pathway thereby ameliorating Con A-induced acute hepatitis, highlighting TAS as a potential drug candidate for treating inflammatory diseases including autoimmune hepatitis.

Immunomodulation Effect of Kaempferia galanga Ethanolic Extract On In Vitro Lymphocyte Cell Proliferation

&lt;p&gt;&lt;em&gt;Kaempferia galanga&lt;/em&gt; L. is a traditional medicine with antitumor properties, as indicated by its immunomodulatory activities. This study aimed to determine the effect of &lt;em&gt;K. galanga&lt;/em&gt; on lymphocyte cell proliferation activity as an indicator of immunomodulatory properties. This study was conducted at the Indonesian Research Center for Veterinary Science (February to April 2018). The immunomodulatory activity of the extract was evaluated with an &lt;em&gt;in vitro&lt;/em&gt; splenocyte proliferation assay. The assay was based on cellular enzymatic synthesis to transform the XTT from formazan tetrazolium as an indicator. The &lt;em&gt;K. galanga&lt;/em&gt; extract was obtained by 96% ethanol extraction. The test was conducted in an aseptic condition, consisted of five treatment groups with three replications each. Three groups of splenocyte cell culture, each with extract concentration of 2.5 µg.ml&lt;sup&gt;-1&lt;/sup&gt;, 25 µg.ml&lt;sup&gt;-1&lt;/sup&gt;, and 250 µg.ml&lt;sup&gt;-1&lt;/sup&gt;, as well as a positive (Concanavalin A/Con A) and negative (cell only) control. The cell suspension (10x10&lt;sup&gt;4 &lt;/sup&gt;cells/ml) was distributed on 96-well plates and cultured following the treatment groups. The same five plates were made for five days of observation and retrieved daily by observing an Elisa reader at 450 nm. The extract of &lt;em&gt;K. galanga &lt;/em&gt;at 2.5 µg.ml&lt;sup&gt;-1&lt;/sup&gt;, 25 µg.ml&lt;sup&gt;-1&lt;/sup&gt;, and 250 µg.ml&lt;sup&gt;-1&lt;/sup&gt; significantly (P &amp;lt;0.05) promoted splenocyte proliferation compared to control. Therefore, it was expected that &lt;em&gt;K. galanga &lt;/em&gt;has a high potential to be used as immunomodulators. Hence, further investigations should be done to clarify the mechanisms of the immunomodulatory effect of &lt;em&gt;K. galanga&lt;/em&gt; as an antitumor &lt;em&gt;in vivo&lt;/em&gt;.&lt;/p&gt;

Uji In Vitro Aktivitas Imunomodulator Minyak Atsiri Serai Dapur (Cymbopogon citratus) terhadap Proliferasi Sel Limfosit Mencit

This study aims to determine the immunomodulatory activity of lemongrass essential oil on the proliferation of mouse lymphocyte cells in vitro. The method used is steam distillation to obtain citronella essential oil. Isolation of lymphocyte cells was obtained from the spleen organ of Swiss-Webster strain mice which was carried out aseptically. The lymphocyte cell proliferation test was carried out using the MTT Assay method and the absorbance was measured using an ELISA reader at 550 nm. The results showed that the lemongrass essential oil contained positive alkaloids, flavonoids, saponins, and steroids/triterpenoids with an essential oil yield of 0.043%. The higher the volatile oil concentration, the higher the Optical Density (OD) value obtained, but the OD increase at a 50 to 100 g/mL concentration was not significantly different. In conclusion, lemongrass essential oil has immunomodulatory activity against lymphocyte cell proliferation in mice tested in vitro.&#x0D; Keywords: Immunomodulator, Essential Oil, Lemongrass (Cymbopogon citratus), Immune System