Life is a continuous process of development. It is beautiful when balanced between passion and duty. The drive of my life is the curiosity to understand the working principle of the natural world and deliver a sincere effort to contribute, at least a little, in this understanding. I am an Immuno-biologist searching the relationship of immunity with disease, development and evolution.
The presence of microglial cells as resident macrophage population in the Central Nervous System ... more The presence of microglial cells as resident macrophage population in the Central Nervous System (CNS) is well documented from the study of repairing of lesions in CNS that varies widely throughout the animal kingdom. The existence of neuroglia cells similar to vertebrate microglia and small mobile phagocytes and hemocytes were documented from ganglia of some invertebrate animal models like leech (H. medicinalis), insects (P. americana and D. melanogaster) and mollusca (M. edulis). Neuronal replacement and migration of immunocompetent cells (macrophage, microglia, ependymal cells etc.) after surgical lesions in CNS of non-mammals (fishes, reptiles and aves) are much restricted to specific neurogenic niches associated to the neural regeneration and migration of cells in invertebrates. Microglial presence is largely restricted in the optic tract of fish and amphibian ganglionic cells because they have a surprising capacity to regenerate their neurons after lesions. Hence the CNS of bo...
Microglial cells are considered to be the chief immunomodulatory cells of the brain. These cells ... more Microglial cells are considered to be the chief immunomodulatory cells of the brain. These cells play a crucial role against various neurodegenerative diseases. When modulated microglia have been shown to exert a potential anti-tumor immune response against brain neoplasms. Although several specific BRMs like IL-2, IFNgamma have been shown to modulate the microglia to get an effective anti-tumor immune response, associated toxicities and detrimental side effects have posed severe limitation in there use particularly for therapeutic purposes. In the present study, attempts have been made to elicit the modulations of microglia cell function and phenotypic expression following specific (IL-2, IFNgamma and a novel nonspecific BRM (corpuscular antigen) in order to determine their anti-tumor property in experimental glioma model. Brain was experimentally induced in young Druckray rats of both sexes with N-N-ethyl nitroso urea (ENU). These ENU treated animals were administered with both sp...
Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic s... more Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic stages and integrate in CNS, are involved in almost all neuroinflammatory conditions. We studied how microglia change their responses through the development and maturation of brain in normal physiological conditions using an ex situ model to delineate their age-specific morpho-functional responsiveness. Rapidly isolated microglia from different age-matched rats were characterized with Iba1 + /CD11b/c + /MHCclassII + , cultured, studied for cell-cycle/proliferative potency, ROS generation and phagocytosis, viability and morphological analysis induced with GMCSF, MCSF, IL-4, IL-6, IL-10, and IFN-γ. The study showed marked differences in cellular properties, stability, and viability of microglia through ontogeny with specific patterns in their studied functions which were coherent with their in situ morphofunctional attributes. Phagocytic behavior showed a notable shift from ROS independence to dependence toward maturation. Perinatal microglia were found persistent in ex situ environment and neonatal microglia qualified as the most potent and versatile responders for morpho-functional variations under cytokine induced conditions. The study identified that microglia from infants were the most stable, adaptive, and better responders, which can perform as an ex situ model system to study microglial biology.
Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic s... more Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic stages and integrate in CNS, are involved in almost all neuroinflammatory conditions. We studied how microglia change their responses through the development and maturation of brain in normal physiological conditions using an ex situ model to delineate their age-specific morpho-functional responsiveness. Rapidly isolated microglia from different age-matched rats were characterized with Iba1 + /CD11b/c + /MHCclassII + , cultured, studied for cell-cycle/proliferative potency, ROS generation and phagocytosis, viability and morphological analysis induced with GMCSF, MCSF, IL-4, IL-6, IL-10, and IFN-γ. The study showed marked differences in cellular properties, stability, and viability of microglia through ontogeny with specific patterns in their studied functions which were coherent with their in situ morphofunctional attributes. Phagocytic behavior showed a notable shift from ROS independence to dependence toward maturation. Perinatal microglia were found persistent in ex situ environment and neonatal microglia qualified as the most potent and versatile responders for morpho-functional variations under cytokine induced conditions. The study identified that microglia from infants were the most stable, adaptive, and better responders, which can perform as an ex situ model system to study microglial biology.
Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic s... more Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic stages and integrate in CNS, are involved in almost all neuroinflammatory conditions. We studied how microglia change their responses through the development and maturation of brain in normal physiological conditions using an ex situ model to delineate their age-specific morpho-functional responsiveness. Rapidly isolated microglia from different age-matched rats were characterized with Iba1 + /CD11b/c + /MHCclassII + , cultured, studied for cell-cycle/proliferative potency, ROS generation and phagocytosis, viability and morphological analysis induced with GMCSF, MCSF, IL-4, IL-6, IL-10, and IFN-γ. The study showed marked differences in cellular properties, stability, and viability of microglia through ontogeny with specific patterns in their studied functions which were coherent with their in situ morphofunctional attributes. Phagocytic behavior showed a notable shift from ROS independence to dependence toward maturation. Perinatal microglia were found persistent in ex situ environment and neonatal microglia qualified as the most potent and versatile responders for morpho-functional variations under cytokine induced conditions. The study identified that microglia from infants were the most stable, adaptive, and better responders, which can perform as an ex situ model system to study microglial biology.
The significant insights into the immunobiology of central nervous system (CNS) and brain tumor h... more The significant insights into the immunobiology of central nervous system (CNS) and brain tumor have opened up the feasibility of applying 'Immunotherapy' as an alternative to the poor prognosis of malignant brain tumor with conventional therapeutic approaches. Though cytokines like IL-2 and IFN-gamma used against glioma showed some favorable results by eliciting Th1 type immune response, a proper immunotherapeutic agent is still to be searched for. Sheep erythrocyte (SRBC), a corpuscular antigen showed a better therapeutic efficacy in terms of enhanced survival and augmentation of cell mediated immunity (CMI) in a glioma model developed by chemical carcinogen ethyl nitrosourea. Histological findings revealed most efficient glioma rejection in SRBC and combination biological response modifier (BRM) treated groups. Simultaneously E-rosetting, cytotoxicity of lymphocytes, phagocytosis and antigen presenting capacity of myeloid cells established the better therapeutic efficacy ...
1 Cellular & Molecular Immunology Lab, Department of Physiology, University College of Medicine, ... more 1 Cellular & Molecular Immunology Lab, Department of Physiology, University College of Medicine, 244 B, AJCBose Road, Kolkata - 700 020. West Bengal, India; 2 Immunohaematology Lab,Department of Haematology, School of Tropical Medicine, CRAvenue, Kolkata 700 ...
Sheep red blood cell (SRBC), a non-specific biological response
modifier that has long been used ... more Sheep red blood cell (SRBC), a non-specific biological response modifier that has long been used as a classical antigen, has been shown to exert an immunomodulatory and anti-tumor activities in experimental animals. The active component of SRBC, which is responsible for such effects, was found to be a cell surface acidic glycoprotein molecule, known as T11 target structure (T11TS). In the present study, T11TS was isolated and purified to homogeneity using a five-step protocol involving isolation of sheep erythrocyte membrane from packed cell volume, 20% ammonium sulfate cut of the crude membrane proteins mixture, immunoaffinity purification using mouse anti-sheep CD58 mAb (L180/1) tagged matrix, preparative gel electrophoresis, and gel electroelution process. Finally, the purity and identity of the proteins were confirmed by the matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analysis. The in silico glycosylation site analysis showed that the extracellular domain contained three N-glycosylation sites (N-12, N-62, and N- 111) and one O-glycosylation site (T-107). However, the experimental analysis negated the presence of O-linked glycan moieties on T11TS. To investigate the role of glycan moieties in the current immunotherapeutic regime, T11TS and its deglycosylated form (dT11TS) were administered intraperitoneally (i.p.) in N-ethyl-N-nitrosoureainduced immune-compromised mice at 0.4 mg/kg body weight. It was observed that both the forms of T11TS could activate the compromised immune status of mice by augmenting immune receptor expression (CD2, CD25, CD8, and CD11b), T-helper 1 shift of cytokine network, enhanced cytotoxicity, and phagocytosis activity. Therefore, the results nullify the active involvement of the N-linked glycan moieties in immunotherapeutic efficacy of T11TS.
The antigen presentation to lymphocytes in brain occurs in two steps. Initially it happens at per... more The antigen presentation to lymphocytes in brain occurs in two steps. Initially it happens at perivascular spaces by perivascular microglia/macrophage population and finally at the site of inflammation deep into brain parenchyma by the resident microglia. But recent evidence challanges the existing notion of involvement of distinct and different cells at these sites. Studies have shown that many of these microglial cells show dendritic cell phenotype in pathogenic and cytokine driven environment. Different subsets of the cell show wide range of myeloid lineage functions indicating a pre-differentiated status of the cell. Monocytic CD34+/B220+ precursor cells have been transformed to microglial cells in vitro and transplantation of these cells show Iba-1 or F4/80 positivity with microglial phenotypes in vivo in adults. Even they can be converted into dendritic cell like forms. The interconvertability among macrophage-microglia-dendritic cells and final effector maturation according to the microenvironmental cues indicates existence of a pre-mature myeloid cell population concerned with antigen presentation and related functions in brain. With the substantial recent observation this article sketches the idea that brain APCs appearing as macrophage/microglia/DC like forms are derivatives of the same stock in response to their position and microenvironment. And also microglia is never any distinct cells, both in neonatal stage and adults.
Free radicals and allied molecules are the potential threats for the cellular components when the... more Free radicals and allied molecules are the potential threats for the cellular components when they are produced in
excess amount and cause different pathophysiological disorders including aging. Contrary to their detrimental effects, these
molecules, in the other hand, can be utilized by the Phagocytic cells to destroy the abnormal cells and cellular components.
Generation of reactive oxygen species (ROS) and nitric oxide (NO) are used as important effector molecules by Phagocytic
macrophage/microglia to eliminate neoplastic cells in glioma bearing rat model. The glycoprotein T11TS/SLFA-3, by
binding with CD2 receptor of macrophage/microglia, induces the generation of these reactive species when applied in three
consecutive doses in glioma bearing animals. The CR3 family receptor CD11b is also correlated with the Phagocytic activity
of the cells. The ‘controlled’ and directed production of ROS and NO by phagocytes induce cell death signals to the glioma
cells and resulted in phagocytic destruction and apoptosis. The death signals generated by the free radicals and associated
molecules resulted in accumulation of p53 proteins in the glioma cells. This oxidative stress induced p53 protein
accumulation in neoplastic cells direct them to die by apoptosis. Therefore, the same oxidative stress causing
pathophysiological problems, are used here to destroy the glioma cells by the macrophage/microglia in the delicate CNS
tissue.
The presence of microglial cells as resident macrophage population in the Central Nervous System ... more The presence of microglial cells as resident macrophage population in the Central Nervous System (CNS) is well documented from the study of repairing of lesions in CNS that varies widely throughout the animal kingdom. The existence of neuroglia cells similar to vertebrate microglia and small mobile phagocytes and hemocytes were documented from ganglia of some invertebrate animal models like leech (H. medicinalis), insects (P. americana and D. melanogaster) and mollusca (M. edulis). Neuronal replacement and migration of immunocompetent cells (macrophage, microglia, ependymal cells etc.) after surgical lesions in CNS of non-mammals (fishes, reptiles and aves) are much restricted to specific neurogenic niches associated to the neural regeneration and migration of cells in invertebrates. Microglial presence is largely restricted in the optic tract of fish and amphibian ganglionic cells because they have a surprising capacity to regenerate their neurons after lesions. Hence the CNS of bo...
Microglial cells are considered to be the chief immunomodulatory cells of the brain. These cells ... more Microglial cells are considered to be the chief immunomodulatory cells of the brain. These cells play a crucial role against various neurodegenerative diseases. When modulated microglia have been shown to exert a potential anti-tumor immune response against brain neoplasms. Although several specific BRMs like IL-2, IFNgamma have been shown to modulate the microglia to get an effective anti-tumor immune response, associated toxicities and detrimental side effects have posed severe limitation in there use particularly for therapeutic purposes. In the present study, attempts have been made to elicit the modulations of microglia cell function and phenotypic expression following specific (IL-2, IFNgamma and a novel nonspecific BRM (corpuscular antigen) in order to determine their anti-tumor property in experimental glioma model. Brain was experimentally induced in young Druckray rats of both sexes with N-N-ethyl nitroso urea (ENU). These ENU treated animals were administered with both sp...
Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic s... more Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic stages and integrate in CNS, are involved in almost all neuroinflammatory conditions. We studied how microglia change their responses through the development and maturation of brain in normal physiological conditions using an ex situ model to delineate their age-specific morpho-functional responsiveness. Rapidly isolated microglia from different age-matched rats were characterized with Iba1 + /CD11b/c + /MHCclassII + , cultured, studied for cell-cycle/proliferative potency, ROS generation and phagocytosis, viability and morphological analysis induced with GMCSF, MCSF, IL-4, IL-6, IL-10, and IFN-γ. The study showed marked differences in cellular properties, stability, and viability of microglia through ontogeny with specific patterns in their studied functions which were coherent with their in situ morphofunctional attributes. Phagocytic behavior showed a notable shift from ROS independence to dependence toward maturation. Perinatal microglia were found persistent in ex situ environment and neonatal microglia qualified as the most potent and versatile responders for morpho-functional variations under cytokine induced conditions. The study identified that microglia from infants were the most stable, adaptive, and better responders, which can perform as an ex situ model system to study microglial biology.
Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic s... more Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic stages and integrate in CNS, are involved in almost all neuroinflammatory conditions. We studied how microglia change their responses through the development and maturation of brain in normal physiological conditions using an ex situ model to delineate their age-specific morpho-functional responsiveness. Rapidly isolated microglia from different age-matched rats were characterized with Iba1 + /CD11b/c + /MHCclassII + , cultured, studied for cell-cycle/proliferative potency, ROS generation and phagocytosis, viability and morphological analysis induced with GMCSF, MCSF, IL-4, IL-6, IL-10, and IFN-γ. The study showed marked differences in cellular properties, stability, and viability of microglia through ontogeny with specific patterns in their studied functions which were coherent with their in situ morphofunctional attributes. Phagocytic behavior showed a notable shift from ROS independence to dependence toward maturation. Perinatal microglia were found persistent in ex situ environment and neonatal microglia qualified as the most potent and versatile responders for morpho-functional variations under cytokine induced conditions. The study identified that microglia from infants were the most stable, adaptive, and better responders, which can perform as an ex situ model system to study microglial biology.
Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic s... more Microglia, myelo-monocytic lineage cells, that enter in the developing brain at early embryonic stages and integrate in CNS, are involved in almost all neuroinflammatory conditions. We studied how microglia change their responses through the development and maturation of brain in normal physiological conditions using an ex situ model to delineate their age-specific morpho-functional responsiveness. Rapidly isolated microglia from different age-matched rats were characterized with Iba1 + /CD11b/c + /MHCclassII + , cultured, studied for cell-cycle/proliferative potency, ROS generation and phagocytosis, viability and morphological analysis induced with GMCSF, MCSF, IL-4, IL-6, IL-10, and IFN-γ. The study showed marked differences in cellular properties, stability, and viability of microglia through ontogeny with specific patterns in their studied functions which were coherent with their in situ morphofunctional attributes. Phagocytic behavior showed a notable shift from ROS independence to dependence toward maturation. Perinatal microglia were found persistent in ex situ environment and neonatal microglia qualified as the most potent and versatile responders for morpho-functional variations under cytokine induced conditions. The study identified that microglia from infants were the most stable, adaptive, and better responders, which can perform as an ex situ model system to study microglial biology.
The significant insights into the immunobiology of central nervous system (CNS) and brain tumor h... more The significant insights into the immunobiology of central nervous system (CNS) and brain tumor have opened up the feasibility of applying 'Immunotherapy' as an alternative to the poor prognosis of malignant brain tumor with conventional therapeutic approaches. Though cytokines like IL-2 and IFN-gamma used against glioma showed some favorable results by eliciting Th1 type immune response, a proper immunotherapeutic agent is still to be searched for. Sheep erythrocyte (SRBC), a corpuscular antigen showed a better therapeutic efficacy in terms of enhanced survival and augmentation of cell mediated immunity (CMI) in a glioma model developed by chemical carcinogen ethyl nitrosourea. Histological findings revealed most efficient glioma rejection in SRBC and combination biological response modifier (BRM) treated groups. Simultaneously E-rosetting, cytotoxicity of lymphocytes, phagocytosis and antigen presenting capacity of myeloid cells established the better therapeutic efficacy ...
1 Cellular & Molecular Immunology Lab, Department of Physiology, University College of Medicine, ... more 1 Cellular & Molecular Immunology Lab, Department of Physiology, University College of Medicine, 244 B, AJCBose Road, Kolkata - 700 020. West Bengal, India; 2 Immunohaematology Lab,Department of Haematology, School of Tropical Medicine, CRAvenue, Kolkata 700 ...
Sheep red blood cell (SRBC), a non-specific biological response
modifier that has long been used ... more Sheep red blood cell (SRBC), a non-specific biological response modifier that has long been used as a classical antigen, has been shown to exert an immunomodulatory and anti-tumor activities in experimental animals. The active component of SRBC, which is responsible for such effects, was found to be a cell surface acidic glycoprotein molecule, known as T11 target structure (T11TS). In the present study, T11TS was isolated and purified to homogeneity using a five-step protocol involving isolation of sheep erythrocyte membrane from packed cell volume, 20% ammonium sulfate cut of the crude membrane proteins mixture, immunoaffinity purification using mouse anti-sheep CD58 mAb (L180/1) tagged matrix, preparative gel electrophoresis, and gel electroelution process. Finally, the purity and identity of the proteins were confirmed by the matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analysis. The in silico glycosylation site analysis showed that the extracellular domain contained three N-glycosylation sites (N-12, N-62, and N- 111) and one O-glycosylation site (T-107). However, the experimental analysis negated the presence of O-linked glycan moieties on T11TS. To investigate the role of glycan moieties in the current immunotherapeutic regime, T11TS and its deglycosylated form (dT11TS) were administered intraperitoneally (i.p.) in N-ethyl-N-nitrosoureainduced immune-compromised mice at 0.4 mg/kg body weight. It was observed that both the forms of T11TS could activate the compromised immune status of mice by augmenting immune receptor expression (CD2, CD25, CD8, and CD11b), T-helper 1 shift of cytokine network, enhanced cytotoxicity, and phagocytosis activity. Therefore, the results nullify the active involvement of the N-linked glycan moieties in immunotherapeutic efficacy of T11TS.
The antigen presentation to lymphocytes in brain occurs in two steps. Initially it happens at per... more The antigen presentation to lymphocytes in brain occurs in two steps. Initially it happens at perivascular spaces by perivascular microglia/macrophage population and finally at the site of inflammation deep into brain parenchyma by the resident microglia. But recent evidence challanges the existing notion of involvement of distinct and different cells at these sites. Studies have shown that many of these microglial cells show dendritic cell phenotype in pathogenic and cytokine driven environment. Different subsets of the cell show wide range of myeloid lineage functions indicating a pre-differentiated status of the cell. Monocytic CD34+/B220+ precursor cells have been transformed to microglial cells in vitro and transplantation of these cells show Iba-1 or F4/80 positivity with microglial phenotypes in vivo in adults. Even they can be converted into dendritic cell like forms. The interconvertability among macrophage-microglia-dendritic cells and final effector maturation according to the microenvironmental cues indicates existence of a pre-mature myeloid cell population concerned with antigen presentation and related functions in brain. With the substantial recent observation this article sketches the idea that brain APCs appearing as macrophage/microglia/DC like forms are derivatives of the same stock in response to their position and microenvironment. And also microglia is never any distinct cells, both in neonatal stage and adults.
Free radicals and allied molecules are the potential threats for the cellular components when the... more Free radicals and allied molecules are the potential threats for the cellular components when they are produced in
excess amount and cause different pathophysiological disorders including aging. Contrary to their detrimental effects, these
molecules, in the other hand, can be utilized by the Phagocytic cells to destroy the abnormal cells and cellular components.
Generation of reactive oxygen species (ROS) and nitric oxide (NO) are used as important effector molecules by Phagocytic
macrophage/microglia to eliminate neoplastic cells in glioma bearing rat model. The glycoprotein T11TS/SLFA-3, by
binding with CD2 receptor of macrophage/microglia, induces the generation of these reactive species when applied in three
consecutive doses in glioma bearing animals. The CR3 family receptor CD11b is also correlated with the Phagocytic activity
of the cells. The ‘controlled’ and directed production of ROS and NO by phagocytes induce cell death signals to the glioma
cells and resulted in phagocytic destruction and apoptosis. The death signals generated by the free radicals and associated
molecules resulted in accumulation of p53 proteins in the glioma cells. This oxidative stress induced p53 protein
accumulation in neoplastic cells direct them to die by apoptosis. Therefore, the same oxidative stress causing
pathophysiological problems, are used here to destroy the glioma cells by the macrophage/microglia in the delicate CNS
tissue.
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Papers by Anirban Ghosh
modifier that has long been used as a classical
antigen, has been shown to exert an immunomodulatory
and anti-tumor activities in experimental animals. The
active component of SRBC, which is responsible for such
effects, was found to be a cell surface acidic glycoprotein
molecule, known as T11 target structure (T11TS). In the
present study, T11TS was isolated and purified to homogeneity
using a five-step protocol involving isolation of
sheep erythrocyte membrane from packed cell volume,
20% ammonium sulfate cut of the crude membrane proteins
mixture, immunoaffinity purification using mouse
anti-sheep CD58 mAb (L180/1) tagged matrix, preparative
gel electrophoresis, and gel electroelution process.
Finally, the purity and identity of the proteins were confirmed
by the matrix-assisted laser desorption/ionization
(MALDI) mass spectrometric analysis. The in silico glycosylation
site analysis showed that the extracellular domain
contained three N-glycosylation sites (N-12, N-62, and N-
111) and one O-glycosylation site (T-107). However, the
experimental analysis negated the presence of O-linked
glycan moieties on T11TS. To investigate the role of
glycan moieties in the current immunotherapeutic regime,
T11TS and its deglycosylated form (dT11TS) were administered
intraperitoneally (i.p.) in N-ethyl-N-nitrosoureainduced
immune-compromised mice at 0.4 mg/kg body
weight. It was observed that both the forms of T11TS
could activate the compromised immune status of mice by
augmenting immune receptor expression (CD2, CD25,
CD8, and CD11b), T-helper 1 shift of cytokine network,
enhanced cytotoxicity, and phagocytosis activity. Therefore,
the results nullify the active involvement of the N-linked glycan moieties in immunotherapeutic efficacy of T11TS.
Keywords : Macrophage, Microglia, Dendritic Cells, Mononuclear phagocytic system (MPS), Differentiation
excess amount and cause different pathophysiological disorders including aging. Contrary to their detrimental effects, these
molecules, in the other hand, can be utilized by the Phagocytic cells to destroy the abnormal cells and cellular components.
Generation of reactive oxygen species (ROS) and nitric oxide (NO) are used as important effector molecules by Phagocytic
macrophage/microglia to eliminate neoplastic cells in glioma bearing rat model. The glycoprotein T11TS/SLFA-3, by
binding with CD2 receptor of macrophage/microglia, induces the generation of these reactive species when applied in three
consecutive doses in glioma bearing animals. The CR3 family receptor CD11b is also correlated with the Phagocytic activity
of the cells. The ‘controlled’ and directed production of ROS and NO by phagocytes induce cell death signals to the glioma
cells and resulted in phagocytic destruction and apoptosis. The death signals generated by the free radicals and associated
molecules resulted in accumulation of p53 proteins in the glioma cells. This oxidative stress induced p53 protein
accumulation in neoplastic cells direct them to die by apoptosis. Therefore, the same oxidative stress causing
pathophysiological problems, are used here to destroy the glioma cells by the macrophage/microglia in the delicate CNS
tissue.
modifier that has long been used as a classical
antigen, has been shown to exert an immunomodulatory
and anti-tumor activities in experimental animals. The
active component of SRBC, which is responsible for such
effects, was found to be a cell surface acidic glycoprotein
molecule, known as T11 target structure (T11TS). In the
present study, T11TS was isolated and purified to homogeneity
using a five-step protocol involving isolation of
sheep erythrocyte membrane from packed cell volume,
20% ammonium sulfate cut of the crude membrane proteins
mixture, immunoaffinity purification using mouse
anti-sheep CD58 mAb (L180/1) tagged matrix, preparative
gel electrophoresis, and gel electroelution process.
Finally, the purity and identity of the proteins were confirmed
by the matrix-assisted laser desorption/ionization
(MALDI) mass spectrometric analysis. The in silico glycosylation
site analysis showed that the extracellular domain
contained three N-glycosylation sites (N-12, N-62, and N-
111) and one O-glycosylation site (T-107). However, the
experimental analysis negated the presence of O-linked
glycan moieties on T11TS. To investigate the role of
glycan moieties in the current immunotherapeutic regime,
T11TS and its deglycosylated form (dT11TS) were administered
intraperitoneally (i.p.) in N-ethyl-N-nitrosoureainduced
immune-compromised mice at 0.4 mg/kg body
weight. It was observed that both the forms of T11TS
could activate the compromised immune status of mice by
augmenting immune receptor expression (CD2, CD25,
CD8, and CD11b), T-helper 1 shift of cytokine network,
enhanced cytotoxicity, and phagocytosis activity. Therefore,
the results nullify the active involvement of the N-linked glycan moieties in immunotherapeutic efficacy of T11TS.
Keywords : Macrophage, Microglia, Dendritic Cells, Mononuclear phagocytic system (MPS), Differentiation
excess amount and cause different pathophysiological disorders including aging. Contrary to their detrimental effects, these
molecules, in the other hand, can be utilized by the Phagocytic cells to destroy the abnormal cells and cellular components.
Generation of reactive oxygen species (ROS) and nitric oxide (NO) are used as important effector molecules by Phagocytic
macrophage/microglia to eliminate neoplastic cells in glioma bearing rat model. The glycoprotein T11TS/SLFA-3, by
binding with CD2 receptor of macrophage/microglia, induces the generation of these reactive species when applied in three
consecutive doses in glioma bearing animals. The CR3 family receptor CD11b is also correlated with the Phagocytic activity
of the cells. The ‘controlled’ and directed production of ROS and NO by phagocytes induce cell death signals to the glioma
cells and resulted in phagocytic destruction and apoptosis. The death signals generated by the free radicals and associated
molecules resulted in accumulation of p53 proteins in the glioma cells. This oxidative stress induced p53 protein
accumulation in neoplastic cells direct them to die by apoptosis. Therefore, the same oxidative stress causing
pathophysiological problems, are used here to destroy the glioma cells by the macrophage/microglia in the delicate CNS
tissue.