CD Markers Research Articles

Real world outcomes with Ibrutinib monotherapy in Chronic Lymphocytic Leukemia: a single center experience

Introduction. The advent of Bruton’s tyrosine kinase (BTK) inhibitors brought about a paradigm shift in the management of chronic lymphocytic leukemia (CLL), by offering a well-tolerated chemotherapy-free approach. Here, we share the experience with ibrutinib of a major Romanian regional cancer center. Methods. We screened patients treated for CLL in our center over 6 years (2017- 2022) and included those who were treated with ibrutinib either in the first line of therapy or in subsequent lines. Results. We enrolled 61 patients, 40 with treatment-naïve (TN) CLL and 21 with relapsed/refractory (R/R) CLL, with a median age at treatment initiation of 65 years. Concerning the prognostic-predictive workup, IgHV mutational status was available for 78.7% of the patients, TP53 sequencing for 82%, assessment of 17p deletion for 82%, and CD38 marker analysis was performed for 70.5%. With a median follow-up period of 55 months, the overall response rate (ORR) was 90.2%, with a median progression-free survival (PFS) of 33 months and a median overall survival (OS) that has not been reached. In our cohort, albeit non-significant statistically, patients with TP53 mutation had a shorter OS and those with mutated IgHV, a shorter PFS. Rai 3-4 and Binet C stages at diagnosis were associated with a shorter PFS, but not OS. In our cohort, the correlation between survival and high Cumulative Illness Rating Scale (CIRS) index was not statistically significant. Ibrutinib was generally well tolerated in our cohort, as only 14.8% of our patients discontinued treatment due to adverse effects. Conclusion. Our study suggests that ibrutinib is a valid therapeutic option for TN or R/R CLL patients, with a high ORR and a good safety profile.

An early precursor CD8 T cell that adapts to acute or chronic viral infection.

This study examines the origin and differentiation of stem-like CD8+ T cells that are essential for sustained T cell immunity in chronic viral infections and cancer and also play a key role in PD-1 directed immunotherapy1-10. These PD-1+ TCF-1+ TOX+ stem-like CD8+ T cells, also referred to as precursors of exhausted T cells8,9, have a distinct program that allows them to adapt to chronic antigen stimulation. Using the mouse model of chronic LCMV infection we found that virus specific stem-like CD8+ T cells are generated early (day 5) during chronic infection suggesting that this crucial fate commitment occurs irrespective of infection outcome. Indeed, we found that nearly identical populations of stem-like CD8+ T cells were generated early during acute or chronic LCMV infection, and that antigen was essential for maintaining the stem-like phenotype. We next performed reciprocal adoptive transfer experiments to determine the fate of these early stem-like CD8+ T cells after viral clearance versus persistence. Following transfer of day 5 stem-like CD8+ T cells from chronically infected into acutely infected mice, these cells downregulated canonical markers of the chronic stem-like CD8+ T cells and expressed markers (CD127 and CD62L) associated with central memory CD8+ T cells. Reciprocally, when day 5 stem-like cells from acutely infected mice were transferred into chronically infected mice these CD8+ T cells functioned like chronic resource cells and responded effectively to PD-1 therapy. These findings highlight the ability of these early PD-1+ TCF-1+ TOX+ stem-like CD8+ T cells to adapt their differentiation trajectory to either an acute or chronic viral infection. Most importantly, our study shows that the host is prepared a priori to deal with a potential chronic infection.

Association of stromal tumor-infiltrating lymphocytes with clinicopathological parameters in endometrial cancer.

Endometrial cancer (EC) ranks as one of the most prevalent gynecological malignancies globally. The presence and role of stromal tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment have garnered interest due to their prognostic and therapeutic potential. This study aimed to evaluate the association between stromal TILs and various clinicopathological parameters in EC. A prospective study was conducted which included 30 histologically confirmed cases of endometrial carcinoma. Specimens collected from January 2023 to June 2024 were processed for routine histopathological examination and immunohistochemistry for CD3 and CD20 markers. TILs were quantified as per the International Immuno-Oncology Biomarker Working Group guidelines and categorized into low (<20%) and high (≥20%) TILs. The study comprised 30 female patients, predominantly aged 51 to 60 years. Most tumors were of the endometrioid subtype (93.3%). High TILs were significantly associated with early tumor stage, lower grade, lesser myometrial invasion, and absence of nodal involvement on univariate analysis and with lower tumor stage and grade on multivariate analysis. No significant association was found between TILs and age, lymphovascular, or perineural invasion. The findings suggest that high TIL infiltration correlates with favorable tumor characteristics, potentially serving as a prognostic marker for early and less aggressive EC. High TILs were associated with better tumor stage, grade, and reduced nodal involvement, indicating their protective role in tumor progression. However, the lack of association with certain parameters calls for further investigation into the functional state of TILs and their interactions within the tumor microenvironment.

A distinct subgroup of AML resembling the APL immunophenotype is associated with DIC.

The complexity of acute myeloid leukemia (AML) is increasingly recognized through the identification of distinct subgroups, including those with an APL-like immunophenotype characterized by the absence of CD34 and HLA-DR expression, which is widely recognized as a representative immunophenotype in acute promyelocytic leukemia (APL). This study sought to understand the clinical, molecular, and prognostic differences between AML patients with and without this phenotype. This study retrospectively analysed 191 de novo non-M3 AML patients and identified 32 patients with the CD34-HLA-DR- phenotype resembling APL-like immunophenotype, considered as the experimental group. Clinical data, including complete blood count, leukemic blasts, coagulation analysis DIC score, and OS, were collected, and immunophenotypic and molecular data were compared between this group and a control group of patients without this immunophenotype. Patients with the CD34-HLA-DR- immunophenotype in the AML cohort had a significantly greater risk of developing disseminated intravascular coagulation (DIC) than did patients in the control group. Additionally, a lower rate of expression of immunophenotypic clusters of differentiation (CD markers) associated with poor prognosis was observed in the CD34-HLA-DR- group. At the molecular level, an increased frequency of nucleophosmin 1 (NPM1) mutations and increased expression of the Wilms' tumor 1 (WT1) gene were noted in this subgroup. However, contrary to patients with an expected favourable prognosis, patients in the favourable risk group with the CD34-HLA-DR- immunophenotype had significantly shorter overall survival than did patients in the control group. The findings highlight the patients exhibiting the CD34-HLA-DR- immunophenotype as a unique AML subgroup with specific clinical and molecular traits, notably a predisposition to DIC, which affecting prognosis. This finding has implications for risk stratification and potential targeted therapies for AML management.

The expressions of myeloid differentiation and non-lineage specific differentiation antigens among FAB subtypes of acute myeloid leukemia in Iraqi patients

Background. Acute myeloid leukemia (AML) is a complex malignant disorder showing various subtypes with myeloid differentiation and non-lineage specific differentiation antigens (CD markers) expressions. Aim. The aim of this study is to evaluate the expression of myeloid and lymphoid antigens in relation to different French‑American‑British classification (FAB subtypes). Method. Three hundred and forty-four acute myeloid leukemia patients diagnosed based on blast percentage underwent bone marrow aspirate or whole blood collection, and flow cytometry was used to identify blast cells. Results. The expressions of myeloid differentiation antigens in AML patients were commonly observed; CD33 (73.3%), CD13 (68.3%), CD117 (67.4%), CD64 (54.6%), and cMPO (52.9%) in AML cases, with a significant difference in positivity (p-value &lt; 0.05) among FAB classes (M0, M1, M2, M3, M4, M5, and M7). The expression of non-myeloid-associated antigen HLA-DR (34.01%) and CD34 (42.15%) showed high prevalence among FAB subtypes of AML. Regarding CD36 (13.3%), CD35 (13.08%), IREM2 (9.9%), and CD123 (9.01%), the expression was lower than HLA-DR and CD34, with no expression of CD38 in AML patients. A significant difference in expression and distribution of CD markers among AML subclasses was found (p&lt;0.05). Conclusion. A large number of AML cases showed CD33 (73.3%), CD13 (68.3%), and CD117 (67.4%) expressions; these markers give two scores in diagnosis and highly distribution in AML subtype. Although no other leukemia subtypes were found to be linked to AML instances, AML blasts exhibited abnormal lymphoid characteristics, whereby the most prevailing lymphoid marker that was aberrantly expressed in AML was CD7. T-cell markers were more prevalent than B-cell markers.

Molecular Imaging of Tumor-Infiltrating Lymphocytes in Living Animals Using a Novel mCD3 Fibronectin Scaffold.

The interaction between cancer cells and immune cells in the tumor microenvironment (TME) plays a crucial role in determining tumor growth, metastasis, and response to treatment. Tumor-infiltrating lymphocytes (TILs) in TME could be a predictive marker for treatment response in various therapeutic interventions, including chemotherapy and immunotherapy. Thus, imaging the tumor immune microenvironment is important for selecting the optimal treatment strategies in cancer therapy. The CD3 protein represents a promising target for diagnostic imaging of TILs in vivo to assess the immune state of the TME. Although many anti-CD3 antibodies have been explored for this application, the nonspecific immune activation by these antibodies limits their applications. To overcome this issue, we engineered a novel fibronectin III domain (FN3) protein binder (mCD3-FN3;11.8 kDa) against mouse CD3 antigen protein using a yeast display library to image TILs homing in vivo into the TME. We performed in vitro and in vivo assays to test the mCD3-FN3 binder purity as well as in vivo targetability in mouse models of syngeneic tumors. We used near-infrared 800 dye conjugated with mCD3-FN3 (IR800-mCD3-FN3) for in vivo tracking of TILs via optical imaging. We used three different syngeneic tumors in mice (mCD3+ EL4 tumor in C57BL/6 mice, mCD3- CT26 colon tumor, and mCD3- 4T1 breast tumor in BALB/c mice) for imaging TILs in vivo. C57BL/6 mice bearing EL4 tumors were separated into two groups (blocking [Blk] and nonblocking [Nblk]; n = 3 per group) and used for in vivo imaging. Blocking groups received 200 μg of unlabeled mCD3-FN3 2 h prior to the administration of IR800-mCD3-FN3 binder. Each mouse was administered with 25 μg of the IR800-mCD3-FN3 binder and tracked using an IVIS optical imaging system over time. C57BL/6/EL4 mice were imaged at 4 and 24 h post injection of the IR800-mCD3-FN3 binder, and mouse organs were collected at 24 h after final imaging and used for ex vivo histological imaging. In CT26 and 4T1 tumor models, TILs in TME were imaged 4, 24, and 48 h after binder injection. The NIR imaging of EL4 tumors showed that IR800-mCD3-FN3 can detect both TILs within the tumor and the tumor cells with a high signal-to-background ratio 24 h after initial binder injection with a total radiant efficiency (mean TRE ± SD) of 6.5 × 1010 ± 1.5 × 1010 [photons/s]/[μW/cm2]. The animals received preinjection of unlabeled mCD3-FN3(Blk) prior to IR800-mCD3-FN3 binder administration and showed a significant level of fluorescence signal reduction (mean TRE ± SD: 1.6 × 1010 ± 4.1 × 109) in the tumor when compared to the EL4-Nblk tumors (p = 0.006). The mouse group with CT26 and 4T1 tumors where the probe can only bind to TILs within the tumor showed a specific imaging signal (mean TRE ± SD) of 1.1 × 1011 ± 5.2 × 1010 and 9.5 × 1010 ± 4.6 × 1010, respectively, at 48 h p.i. For these groups, the ex vivo tumor-to-muscle ratios were 20- and 27-fold for CT26 and 4T1 tumors, respectively. These results clearly demonstrate the in vivo binding ability of the mCD3-FN3 binder to mCD3 marker expressed by T cells in the TME. The ex vivo histological analysis of tumors, and the organs of animals with EL4 tumors, and TILs imaging of CT26, and 4T1 tumors (at 48 p.i.) confirmed that the IR800-mCD3-FN3 probe was able to specifically bind to CD3 markers expressed by the T cells. In summary, both in vitro and in vivo data indicated that the engineered mCD3-FN3 binder by this study is a promising ligand for diagnostic imaging of tumors in vivo for the assessment of mCD3 expressing TILs in the TME. This can be used as a prognostic marker in evaluating tumor response to therapeutic intervention as well as a diagnostic marker in imaging tumor response to immune checkpoint blockade cancer therapies.

Polycytotoxic T cells mediate antimicrobial activity against intracellular Mycobacterium tuberculosis.

Protection against infections with intracellular bacteria requires the interaction of macrophages and T-lymphocytes, including CD8+ T cells. Recently, the expression of natural killer cell receptors NKG2A and NKG2C was introduced as markers of CD8+ T-cell subsets. The goal of this study was to functionally characterize human NKG2A and NKG2C-expressing T cells using the major pathogen Mycobacterium tuberculosis (Mtb) as a model organism. Sorted NKG2 populations were analyzed for their capacity to proliferate and degranulate and their intracellular expression of cytotoxic molecules. Cytokine release and the effect on bacterial growth were assessed after coculture of NKG2 populations with Mtb-infected macrophages. NKG2A+ T cells released higher levels of IFN-γ and IL-10, whereas NKG2C+ T cells released higher levels of IL-2, contained the greatest reservoir of intracellular granzyme B and showed a remarkable constitutive level of degranulation. Both subsets inhibited the intracellular growth of Mtb more efficiently than NKG2-negative CD8+ T cells. Antimicrobial activity of NKG2+ T cells was not associated with the release of cytokines or cytotoxic molecules. However, the frequency of polycytotoxic T cells (P-CTL), defined as CD8+ T cells co-expressing granzyme B, perforin, and granulysin, positively correlated with the ability of NKG2-expressing T cells to control Mtb-growth in macrophages. Our results highlight the potential of NKG2-expressing P-CTL to trigger the antibacterial activity of human macrophages. Targeting this population by preventive or therapeutic immune interventions could provide a novel strategy to combat severe infectious diseases such as tuberculosis.

Arginine vasopressin (AVP) treatment increases the expression of inhibitory immune checkpoint molecules in monocyte-derived dendritic cells.

Arginine vasopressin (AVP) has disparate impacts on immune responses by divergent receptors on cells including DCs. This study was conducted with the aim of investigating the impact of AVP on the maturation and expression of the inhibitory immune checkpoint molecules in tolerogenic monocyte-derived DCs. CD14 marker was used to separate monocytes from peripheral blood mononuclear cells (PBMCs) by MACS method. To differentiate monocytes from DCs, we utilized GM-CSF and IL-4 cytokines. Tolerogenic DCs were generated using vitamin D3 and dexamethasone. We added LPS and AVP to the culture medium on day 6 after incubation of DCs at 37°C. Finally, we assessed the surface molecules by flow cytometry and used real-time PCR to evaluate the expression of genes related to the inhibitory immune checkpoints. Based on the obtained data, AVP increased the expression of CD11c (P ≤ 0.0001), HLA-DR (P ≤ 0.01), and CD86 (P ≤ 0.01) in AVP-mDCs. Also, the expression of all the immune checkpoint genes including CTLA-4 (P ≤ 0.001), BTLA (P ≤ 0.001), PDL-1 (P ≤ 0.05), B7H7 (P ≤ 0.001), LAG3 (P ≤ 0.01), and VISTA (P ≤ 0.001) in AVP-mDCs was increased in comparison to the control group. Vasopressin caused the generation of mature and tolerogenic DCs. Our data may help to consider AVP-mDCs to take part in autoimmune disease therapy, transplanted tissue rejection impedance, and allergies.

Immunoarchitectural Pattern and Its Potential Prognostic Value in Mucoepidermoid Carcinoma.

To define tumor immunoarchitectural patterns (IPs) and characterize the immune profile in salivary gland mucoepidermoid carcinoma (MEC) toward assessing MEC prognostic significance and implications for immunotherapy. This study analyzed 41 MEC cases, evaluating the tumor IPs and tumor-infiltrating lymphocyte (TIL) parameters by using whole-slide imaging and AI-assisted assessment. Immunohistochemistry of CD3 and CD8 markers was performed to assess key lymphocyte subpopulations. Immune-rich (I-rich) tumors were characterized by high TIL density and were associated with aggressive tumor behavior, particularly in histological high-grade MEC, with a significant increase in TIL density observed in the inner invasive margin compartment (p < 0.05). I-rich tumor cases were linked to poorer disease-free survival (DFS) (Breslow = 4.686, p = 0.03). Aggressive MEC cases displayed a highly heterogeneous TIL profile, however, no TIL patterns showed a significant impact on DFS (p > 0.05). Despite the high immunogenicity suggested by the abundance of T lymphocytes, the protective role of CD8+ T lymphocytes was not prominent in MEC. I-rich tumors appeared more aggressive and unfavorable MEC cases exhibited high heterogeneity in their TIL profiles. Interrogating the role of TILs in the inner invasive tumor margin may offer a new mechanistic understanding for future immunotherapy discovery.

CD3+ Tumor-Infiltrating Lymphocytes in Breast Carcinoma: Their Prognostic Role and Association With Clinical Parameters.

Background Breast carcinoma cases are rising steadily and represent a major causeof mortality and morbidity in India. In response to breast carcinoma, the immune system is activated, resulting in lymphocyteinfiltration in and around the tumor nests. This interaction between the tumor and immune system is the basisfor studyingtumor-infiltrating lymphocytes (TILs). Despite studies on TILs in breast carcinoma, the role of CD3+TILsin predicting patient outcomes remains underexplored. This study aimed to evaluate TILsin breast carcinoma tissues by analyzing CD3 expression through immunohistochemistry (IHC) and examining its association with various prognostic factors, such as histological grade, clinical stage, lymph node status, and hormone receptor status. Methods A study was conducted on 45 breast carcinoma cases, documenting various clinicopathological parameters. IHCstaining was performed for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor (HER2-neu), and CD3 markers on tissue samples. Both intra-tumoral and stromal CD3 TILswere quantified and analyzed in relation to clinicopathological prognostic factors. Results In all 45 breast carcinoma cases, intra-tumoral and stromal CD3 expression was assessed. High stromal CD3 expression is documented in the triple-negative breast tumor. CD3 intra-tumoral expression correlated significantly with tumor size (p < 0.035), ER status (p < 0.036), and PR status (p < 0.036). CD3 stromal expression showed no correlation with any of the prognostic parameters. Conclusion The current study found a strong correlation between CD3 intra-tumoral expression with tumor size, ERand PRstatus. The findings imply that CD3 may be a significant factor in breast carcinoma prognosis. There were more stromal CD3 TILsin triple-negative breast carcinoma (TNBC) cases. TILsought to be examined for their capacity as novel prognostic and predictive indicators, especially in cases of invasive breast cancer, such as triple-negative breast carcinoma.

Opposing effects of mycotoxins alternariol and deoxynivalenol on the immunomodulatory effects of oxaliplatin and triapine.

Mycotoxin occurrence in food worldwide is estimated to increase due to climate change. Moreover, studies on how these food contaminants interfere with medications and especially anticancer therapies are rare. With the rise of anticancer immunotherapies, particularly mycotoxins with immunomodulatory activity, such as alternariol (AOH) or deoxynivalenol (DON), are of great concern. Both mycotoxins interfere with the pro-inflammatory nuclear factor kappa B (NF-κB) pathway in myeloid cells. This pathway not only plays an important role in the anticancer immune response but also inflammatory side effects induced by chemotherapeutic drugs. Consequently, the aim of this study was to investigate possible beneficial or detrimental immunomodulatory interactions between these mycotoxins and anticancer drugs. To assess the combined influence of mycotoxins and anticancer therapies on immune cell stimulation, THP-1 NF-κB reporter cells were utilized as monocytes as well as differentiated and polarized macrophages. Parameters for activation (NF-κB activity and protein expression), differentiation (CD14 and CD71 surface marker expression) and polarization (interleukin 10 (IL10), interleukin 8 (CXCL8), tumor necrosis factor α (TNF), prostaglandin-endoperoxide synthase 2 expression and CXCL8 secretion) were assessed upon combinatory treatment. Both mycotoxins affected the immunostimulatory effects of the pre-selected anticancer drugs oxaliplatin and triapine, although in opposing directions. While AOH generally suppressed a drug-induced activation and increased anti-inflammatory IL10 levels, DON potentiated activation and pro-inflammatory markers, such as CXCL8 and TNF in immune cells. In conclusion, AOH and DON have the potential to alter the immunological effects of anticancer therapies, which should be considered during therapy as well as in their future risk assessment.

Adipose stem-cell-derived microvesicles ameliorate long-term bladder ischemia-induced bladder underactivity.

The mechanism for long-term hypoxia/ischemia induced bladder underactivity is uncertain. It requires an effectively therapeutic treatment. Therefore, we determined the pathophysiologic mechanisms of long-term bilateral partial iliac arterial occlusion (BPAO)-induced bladder underactivity and explored the therapeutic potential of adipose-derived stem cells (ADSCs) and ADSC-derived microvesicles (MVs) on BPAO-induced bladder dysfunction. The study included four groups: sham, BPAO, BPAO+ADSCs, and BPAO+ADSC-MVs. ADSCs or ADSC-MVs were isolated, characterized with specific CD markers and injected through the femoral artery to the rat bladders. Real-time laser speckle contrast imaging evaluated bladder microcirculation after BPAO. The transcystometrogram, pelvic nerve activity, bladder histology, immunohistochemistry, and lipid peroxidation assays were conducted after 4-week BPAO induction. The molecular mechanisms of bladder expression of purinergic P2X2/P2X3 and cholinergic M2/M3 receptors for regulating bladder contractility, nerve growth factor (NGF) for nerve injury repair, and collagen-1 for fibrosis were evaluated. Long-term BPAO significantly reduced bladder microcirculation, prolonged the intercontraction interval, decreased voiding volume, increased residual urine volume, lengthened phase 1 contraction, shortened phase 2 contraction, increased leukocytes and CD68 infiltration, increased malondialdehyde levels, and decreased levels of P2X3 and M3 receptors. ADSC-MVs were more efficient than ADSCs in improving BPAO induced parameters, recovering P2X3 and M3 receptors, increasing NGF expression, and decreasing collagen-1 expression in the bladder. ADSC-derived MVs were better than ADSCs to improve long-term BPAO-induced detrusor underactivity, bladder ischemia, and oxidative stress. ADSC-MVs through the therapeutic action of ameliorating inflammation, improving purinergic/cholinergic signaling and neuronal regeneration, and decreasing fibrosis improved BPAO-induced bladder underactivity.

Analysis of the presence of natural killer cell subpopulations in preterm human milk: A first approach

Several immune cell populations are transferred to the newborn through breast milk, including natural killer (NK) cells, which are critical for innate defense and regulation of the immune response, especially in preterm infants. The aim of this study was to analyze the presence of NK cell subpopulations in different types of preterm breast milk. The study quantified the presence of NK cell subpopulations by flow cytometry using the relative expression of CD56 and CD16 markers in colostrum, transitional and mature milk samples from preterm mothers. Flow cytometry analysis revealed the presence of five NK cell subpopulations, but unlike those reported in peripheral blood, CD56dimCD16+ and CD56-CD16+ populations are predominantly present in preterm milk, only the CD56brightCD16dim population is increased in mature milk. Analysis of NK cell subpopulations in preterm milk revealed a pattern of NK cell presence in preterm breast milk with predominantly cytotoxic phenotypes in relation to CD16 marker expression.

Implications of glioblastoma-derived exosomes in modifying the immune system: state-of-the-art and challenges.

Glioblastoma is a brain cancer with a poor prognosis. Failure of classical chemotherapy and surgical treatments indicates that new therapeutic approaches are needed. Among cell-free options, exosomes are versatile extracellular vesicles (EVs) that carry important cargo across barriers suchas the blood-brain barrier (BBB) to their target cells. Thismakes exosomes an interesting option for the treatment ofglioblastoma. Moreover, exosomes can comprise many therapeutic cargos, including lipids, proteins, and nucleic acids,sampled from special intercellular compartments of their origin cell. Cells exposed to various immunomodulatory stimulican generate exosomes enriched in specific therapeutic molecules. Notably, the secretion of exosomes could modify theimmune response in innate and adaptive immune systems. For instance, glioblastoma-associated exosomes (GBex) uptake bymacrophages could influence macrophage dynamics (e.g., shifting CD markers expression). Expression of critical immunoregulatory proteins such as cytotoxic T-lymphocyte antigen-1 (CTLA1) and programmed death-1 (PD-1) on GBex indicates the direct crosstalk of these nano-size vesicles with the immune system. The present study reviews the role of exosomes in immune system cells, including Bcells, Tcells, natural killer (NK) cells, and dendritic cells (DCs), as well as novel technologies in the field.

ADGRG1/GPR56 As a Marker of CD8+ Tumor-Reactive T Cells in Acute Myeloid Leukemia

ADGRG1/GPR56 As a Marker of CD8+ Tumor-Reactive T Cells in Acute Myeloid Leukemia

Interleukin-12 decorated nanosized semiflexible Immunofilaments enable directed targeting and augmented IFNγ responses of natural killer cells

Immunotherapies are a powerful strategy to treat cancer by modulating the immune system to raise an anti-tumor immune response. A prime example of immunotherapies are cytokines - small immunomodulatory molecules that are widely used to stimulate immune cells. Undirected administration of cytokines, however, can cause severe side effects, preventing the use of potent cytokines, such as Interleukin (IL)-12, which induces IFNγ responses by cytotoxic effector lymphocytes, including NK cells. Biomaterials, like nanoparticles, can encapsulate IL-12 and accumulate at the tumor site to alleviate side effects. Yet, the released IL-12 might not be directly targeted to extracellular IL-12 receptors on the specific effector cells, thereby potentially compromising the cytokine's therapeutic efficacy. Here, we develop a polymer-based platform to target NK cells, which we call immunofilaments. Immunofilaments are nanosized linear polymers that present an anti-CD16 antibody and IL-12 effectively to NK cells and lead to synergistic NK cell activation as highlighted by an increase in TNFα and IFNγ production and upregulation of multiple activation markers, including CD25, CD69, and degranulation marker CD107a. NK cell proliferation is enhanced in the presence of both anti-CD16 antibody and IL-12 compared to giving IL-12 separately. Finally, we demonstrate that the IF platform is suitable for in vivo applications, as immunofilaments readily activate human NK cells upon administration to mice. Statement of SignificanceIL-12 is a potent cytokine that stimulates IFNγ responses in NK cells, which supports an anti-tumor immune response. Due to its high potency, the delivery of IL-12 needs to be highly controlled to prevent severe adverse side effects, which can be achieved by using biomaterials. This study shows that nanosized polymers termed Immunofilaments can be used to immobilize IL-12 and effectively target and activate NK cells by co-conjugation of anti-CD16 antibodies. This work is a prime example of careful engineering of innovative biomaterials to improve immunotherapy.

Investigating T Cell Immune Dynamics and IL-6's Duality in a Microfluidic Lung Tumor Model.

Interleukin 6 (IL-6), produced by immune cells, is crucial in promoting T cell trafficking to infection and inflammation sites, influencing various physiological and pathological processes. Concentrations of IL-6 and other cytokines and chemokines can influence T cell differentiation and activation. Understanding the dual faces of IL-6 within the tumor microenvironment is crucial to understanding its role. A flow-based microsystem was designed to investigate CD4+ T cell activation in response to different IL-6 gradients in an under-control 3D culture. The study found that cancer cells' response to varying IL-6 concentrations was dynamic and dose-sensitive, with immune cell migration rates showing sensitivity to the IL-6 gradient. A549 cell expansion increases gradually and time-dependently with 50 ng of IL-6, while Jurkat cell migration follows a time-dependent pattern. However, when a total of 100 ng IL-6 concentration is applied, A549 cells expand rapidly, potentially influencing Jurkat cell migration. Jurkat cell mobility is lower, possibly due to increased A549 cell presence and heightened cell-cell interactions. Different IL-6 concentration gradients can modulate the expression of some CD markers like CD69 and programed cell death protein 1 in CD4+ T cells, suggesting that IL-6 concentration gradients affect immune cell phenotypes. This suggests that IL-6 plays a crucial role in activating T helper cells and may be involved in the later phases of inflammation. Also, the increased levels of IFN-γ and TNF-α highlight IL-6's impact on T cell inflammatory response. This study emphasizes the intricate effects of IL-6 on T cell activation, phenotype, cytokine production, and phenotypic heterogeneity, providing valuable insights into immune response modulation in an experimental setting.

Impact of Dolutegravir plus Lamivudine as First-Line Antiretroviral Treatment on HIV-1 Reservoir and Inflammatory Markers in Peripheral Blood.

To compare the effects of first-line antiretroviral treatment (ART) with dolutegravir plus lamivudine (DTG+3TC) versus DTG plus emtricitabine/tenofovir alafenamide (FTC/TAF) on the evolution of the HIV-1 reservoir and immune activation biomarkers in people with HIV (PWH). DUALITY was a 48-week, single-center, randomized, open-label clinical trial in ART-naïve PWH. Participants were randomized (1:1) to receive ART with DTG+3TC (2DR group) or DTG+FTC/TAF (3DR group). Total and intact proviral HIV-1 DNA, cell-associated RNA in CD4+ T cells, the frequency of HIV-infected CD4+ T cells able to produce p24, plasma soluble inflammatory markers (IL-6, sCD14, TRAIL, IP-10, FABP2, CRP and D-dimer), and activation and exhaustion markers in CD4+ and CD8+ T cells were longitudinally determined. Forty-four participants (22 per study arm) were enrolled. Baseline mean (SD) log10 plasma viral load (pVL) and CD4+ T cell counts were 4.4 (0.7) copies/mL and 493 (221) cells/mm3, respectively. All participants completing the study (2DR n=20; 3DR n=21) had pVL <50 copies/mL at week 48, except for one in the 2DR group who was resuppressed after treating syphilis. Changes from baseline to week 48 in all reservoir parameters or in levels of soluble inflammatory biomarkers and activated or exhausted CD4+ and CD8+ T cells were similar between 2DR and 3DR groups. First-line ART with DTG+3TC resulted in a similar reduction of HIV-1 persistence parameters in peripheral blood, and comparable changes in immune-associated soluble and T-cell markers compared with DTG+FTC/TAF. These findings support recommendation of DTG/3TC among preferred options for first-line ART in PWH.

Diagnosis of canine B-cell chronic lymphoid leukemia with a CD21 negative phenotype using the LT21 clone CD21 antibody in flow cytometry: a case report

BackgroundChronic lymphoid leukemia (CLL) is a hematological disorder characterized by the clonal expansion of small mature lymphocytes that accumulate in the blood and bone marrow. CLL can arise from B-, T-, or natural killer cell clones. The cytological evaluation of blood smears is often the simplest and least invasive method for diagnosing lymphoid leukemia. Immunophenotyping is used to further subclassify the type of lymphoid leukemia.Case presentationA 15-year-old, 4.4-kg spayed female Shih Tzu was presented to the veterinary medical teaching hospital of Kangwon National University. Despite having a normal appetite and activity level, cervical and inguinal lymph node enlargement was noted on physical examination. Complete blood count revealed severe leukocytosis, severe lymphocytosis, and monocytosis. Splenomegaly, hepatomegaly, and lymph node enlargement were detected on radiographic and ultrasonographic examination. Immunophenotyping was performed using peripheral blood mononuclear cells (PBMCs). The majority of lymphocytes exhibited the following profiles: CD3−CD79a− (97.5%), CD4−CD8− (98.6%), CD21−CD79a− (98.4%), CD34− (0.1%), CD45+ (99.6%), major histocompatibility complex class II+ (99.5%), and CD14− (0.5%). Based on the immunophenotyping results, possible differentials considered included the following: the majority of lymphocytes may be natural killer (NK) cell clones, plasma cell clones, or show aberrant expression or loss of CD21 marker due to the neoplastic nature of the cells. Further flow cytometry was performed using antibodies against CD3, CD5, CD94, and granzyme B. The combined results indicated that the predominant lymphocyte subset in the PBMCs was CD3−CD5−CD21−CD94−granzyme B−. To confirm monoclonality and exclude the aberrant loss of CD markers, a polymerase chain reaction for antigen receptor rearrangement (PARR) assay was conducted. The PARR assay, using DNA from blood and lymph node samples, showed B-cell monoclonality. Immunocytochemistry using PBMCs showed that the plasma cell marker Multiple Myeloma Oncogene 1 (MUM1) was not expressed. Therefore, the diagnosis was confirmed to be B-cell CLL.ConclusionImmunophenotyping can help subclassify the type of lymphoid leukemia; however, as tumor cells can show aberrant expression or loss of the CD21 marker, combining immunophenotyping with the PARR assay could yield a more accurate diagnosis.

The impact of prebiotic, probiotic, and synbiotic supplements on CD4, CD8 counts and inflammatory markers in HIV patients: A systematic review and meta-analysis

The impact of prebiotic, probiotic, and synbiotic supplements on CD4, CD8 counts and inflammatory markers in HIV patients: A systematic review and meta-analysis