This action might not be possible to undo. Are you sure you want to continue?
• The Epstein-Barr virus (EBV), also called human herpesvirus 4 (HHV-4) • EBV is named after Anthony Epstein and Yvonne Barr, who together with Bert Achong discovered the virus in 1964. • It is a member of the herpesvirus family and one of the most common human viruses
• It occur in late adolescence or young adulthood, and primary infection with EBV occurs in childhood and is usually asymptomatic. • is transmitted by close human contact, frequently with the saliva during kissing
• Causes by infectious mononucleosis(glandular fever), a benign, self-limited lymphoproliferative disorder. • Associated with the development of a number of neoplasms, like Burkitt Lymphomas and nasopharyngeal carcinoma.
INFECTIOUS MONONUCLEOSIS • known as the kissing disease comes from oral distribution known as mono. . Mono is a sickness that cases extreme fatigue and general feeling of sickness usually lasting anywhere from 2 to 6 weeks.
EBV INFECTION: Oropharynx Viral Ingestion B-Cells Infectious Mononucleosis •Lymphadenitis • Splenitis • Hepatitis • Pneumonitis • Menigitis •Encephalitis Burkitt Lymphoma .
malaise • Lymphoma • Lymphadenopathy (disorder of Lymph nodes or vessels) • Hepatitis • Pneumonitis • Menigitis •Encephalitis .Classical Symptoms of Epstein Barr Virus • Fever • Sore Throat • Lymphadenitis (Swollen Lymph glands) •splenomegaly Atypical Symptoms of Epstein Barr Virus • Extreme fatigue.
A positive heterophile antibody reaction (monospot test) 3. Lymphocytosis with the characteristic atypical lymphocytes in the peripheral blood. Specific anti-bodies for EBV antigens . 2.DIAGNOSIS OF INFECTIOUS MONONUCLEOSIS DEPENDS ON: 1.
Sarmiento .EPSTEIN BARR VIRUS IN BONE MARROW OF RHEUMATOID ARTHRITIS PATIENTS PREDICTS RESPONSE TO RITUXIMAB TREATMENT Karla Tricia C.
HSV-2 • Polyoma virus .VIRUSES INVOLVED IN RHEUMATOID ARTHRITIS • EBV • Parvovirus B19 • CMV • HSV-1.
increased number of EBV-infected B cells and high prevalence of antibodies to different EBV antigens EBV infection may transform infected B cells into antibody-secreting plasma cells . Sequence homology between gp110 of EBV and the shared epitope (SE) in the MHC Class II gene could be a link between RA and EBV EBV-infected RA patients have a diminished cytotoxic Tcell response to EBV as compared with non-RA controls.
HOW VIRUSES MAY CONTRIBUTE TO ARTHRITIS • RA patients often stand on immunosuppressive treatment that may expose them to an increased risk of viral infection • immunosuppressive treatment that alters immune responses to pre-existing viral infections could theoretically influence response to treatment .
• To determine the presence of parvovirus B19 and EBV before and 3 months after immunosuppressive treatment with rituximab
• To determine the importance of specific viral infections for response to RTX
35 participants ◦ 33 on MTX ◦ 1 AZA ◦ 1 chlorambucil ◦ 34 non-responders to previous anti-TNF-α tx
10 ml of heparinized bone marrow aspirates were collected from crista iliaca peripheral blood obtained by venipuncture from the cubital vein and placed in a sterile heparinized vacuum container
PARAMETER EBV (+) (n=15) Parvo B19 (+) (n=8) Virus (-) (n=12) Age.9 8 (67) 6. n (%) Anti-CCP median. mg/week Chlorambucil AZA HLA-DRB1 carriers. mean.689 .21 12 0 0 1/12 (8) 15.1 14/1 10.50 7 0 1 3/8 (3. median DMARD tx MTX. years Gender. mean. female/male Dse duration.90 14 1 0 6/13 (46) 24.185 54. U/ml 61. n (%) DAS-28 at baseline.058 57.75) 16.3 7/1 17. yrs RF positive at baseline.8 8 (100) 5.7 13 (87) 5.2 9/3 13.
yrs RF positive at baseline.9 Virus (-) (n=12) 13 (87) 5.PARAMETER Age. years Gender.50 7 0 1 3/8 (3.3 7/1 17. female/male Dse duration.8 8 (100) 5. U/ml . mean. median 61.2 9/3 13.90 14 1 0 6/13 (46) 24058 8 (67) 6. mg/week Chlorambucil AZA HLA-DRB1 carriers. n (%) Anti-CCP median. n (%) DAS-28 at baseline.75) 16185 54.21 12 0 0 1/12 (8) 15689 DMARD tx MTX. mean.1 14/1 10.7 EBV (+) (n=15) Parvo B19 (+) (n=8) 57.
• EBV and parvovirus B19 are frequently detected in bone marrow of RA px .
• EBV positivity correlates to a better clinical response to RTX therapy .
PARAMETER Age.50 4. mean. n (%) DAS-28 at baseline.3 DMARD tx MTX.5%) 7 0 1 54. U/ml 6/13 (46) 24058 3/8 (3.3 7/1 17. n (%) Anti-CCP median. n (%) at 6 mos followup.2 9/3 13.08 1 px (12.86 5 px (42%) 12 0 0 HLA-DRB1 carriers. female/male Dse duration. mg/week Chlorambucil AZA 61.95 12 px (80%) 14 1 0 8 (67) 5 (41) 6.75) 16185 1/12 (8) 15689 . median Change DAS28>1. 90 3. mean.8 8 (100) 2 (25) 5. median at 6 mos followup. years Gender.9 Virus (-) (n=12) 13 (87) 9 (60) 5.7 EBV (+) (n=15) Parvo B19 (+) (n=8) 57.1 14/1 10.21 4. yrs RF positive at baseline.
1 14/1 10. U/ml 61. n (%) DAS-28 at baseline. n (%) Anti-CCP median.5%) 7 0 1 3/8 (3.689 .2 9/3 13.75) 16. mean. 90 3. years Gender. mean.058 8 (67) 5 (41) 6. yrs RF positive at baseline.9 Virus (-) (n=12) 13 (87) 9 (60) 5.7 EBV (+) (n=15) Parvo B19 (+) (n=8) 57.95 12 px (80%) 14 1 0 6/13 (46) 24. median Change DAS28>1.08 1 px (12.21 4. mg/week Chlorambucil AZA HLA-DRB1 carriers.PARAMETER Age.185 54. n (%) at 6 mos followup.3 7/1 17. female/male Dse duration. median at 6 mos followup.86 5 px (42%) 12 0 0 1/12 (8) 15.3 DMARD tx MTX.50 4.8 8 (100) 2 (25) 5.
• RTX selectively depletes CD20-expressing B cells .
50 4. n (%) Anti-CCP median. mean. median at 6 mos followup.21 4. yrs RF positive at baseline. 90 3.9 Virus (-) (n=12) 13 (87) 9 (60) 5. n (%) at 6 mos followup.8 8 (100) 2 (25) 5. mg/week Chlorambucil AZA HLA-DRB1 carriers.PARAMETER Age. n (%) DAS-28 at baseline. female/male Dse duration.1 14/1 10.7 EBV (+) (n=15) Parvo B19 (+) (n=8) 57.3 7/1 17. mean.08 1 px (12.86 5 px (42%) 12 0 0 1/12 (8) 15689 .75) 16185 54.2 9/3 13.3 DMARD tx MTX. years Gender.5%) 7 0 1 3/8 (3. U/ml 61. median Change DAS28>1.95 12 px (80%) 14 1 0 6/13 (46) 24058 8 (67) 5 (41) 6.
. naive. • A population of CD19+CD95+ (Fas) expressing B cells in the bone marrow was significantly higher at baseline in EBV-positive than EBV-negative patients.• At baseline and at 3 months after treatment. there were no significant differences in the proportions of bone marrow immature. unswitched memory and switched memory B cells between EBV-positive and EBVnegative patients.
• B-cell depletion eradicated all traces of EBV infection in blood and bone marrow at 3 months post-RTX treatment • Carriers of EBV had a significantly better clinical response to B-cell depletion therapy than did patients without EBV infection .
CYTOLYTIC T-CELL RESPONSE AGAINST EPSTEIN-BARR VIRUS IN LUNG CANCER PATIENTS AND HEALTHY SUBJECTS Irene Kei Bariuad .
as compared to age-matched healthy controls and whether any variation in this response could be attributed to senescence. .ABSTRACT Background This study aimed to examine whether EBV seropositive patients with lung cancer have an altered virus-specific CTL response.
ABSTRACT Methods Peripheral blood mononuclear cells from lung cancer patients. age-matched and younger healthy individuals were used to measure EBVspecific CTLs after in vitro amplification with the GLCTLVAML and RYSIFFDYM peptides followed by HLA-multimer staining. .
ABSTRACT Results Lung cancer patients and aged-matched controls had significantly lesser EBVspecific CTL than younger healthy individuals. . Multimer positive populations from either group did not differ with respect to the percentage of multimer positive CTLs and the intensity of multimer binding.
These data warrant the examination of whether young individuals have a more robust antitumor response. as is the case with the antiEBV response .ABSTRACT Conclusions This study provides evidence that patients with lung cancer exhibit an EBV-specific CTL response equivalent to that of age-matched healthy counterparts.
including the tumor itself.INTRODUCTION • Cancer patients present with a compromised immune response of multifactorial origin. • Early stages of tumor growth appear not to elicit systemic immune deficiency and are sometimes associated with antigen-specific tolerance • Generalized immunodeficiency can arise during the late stages of tumor development .
. EBV seropositive patients with lung cancer.INTRODUCTION • This study was scheduled in order to examine whether. have a compromised virusspecific CTL response. • A group of younger healthy individuals was also examined to ascertain whether a possible reduction in the anti-EBV CTL responses of the above patients and age-matched controls could be attributed to senescence. at diagnosis. as compared to agematched healthy controls.
SUBJECTS AND METHODS Patients and controls 1. . PBMC were isolated from whole blood collected at diagnosis from 19 patients with primary lung cancer.
7.(+) SCLC (small cell lung carcinoma) mean age: 67.11. 3 females.0 +/.SUBJECTS AND METHODS Patients and controls • Thirteen. 1 female.4 years.(+) NSCLC (non small cell lung carcinoma) mean age: 66. 10 males • Six.8 +/. 5 males .8 years.
3 males All PBMC were kept frozen till required .SUBJECTS AND METHODS Patients and controls 2.5. PBMC were also collected from 14 age-matched healthy Individuals mean age: 58.0 years. 10 males 3. 4 females.8 years.PBMC were also collected from 7 healthy younger individuals mean age: 26. 4 females.2 +/.7+/.1.
.SUBJECTS AND METHODS Patients and controls • Subjects expressed HLA-A2 and/or -A24 • There were positive for IgG antibodies against the EBV nuclear antigen 3C (EBNA3C).
A2 presented by HLA-A2) and RYSIFFDYM (EBNA3C.A24 presented by HLA-A24) were used. .DETECTION OF EBV-SPECIFIC CTLS • Peptide-specific CTLs were detected using HLAmultimer flow cytometry after a previous step of in vitro amplification of MLPCs with peptides under limiting dilution conditions • Two EBV peptides • GLCTLVAML(BMLF1.
DETECTION OF EBV-SPECIFIC CTLS • Specific multimers labelled with APC and control multimers with PE were used to stain MLPC. only if staining with the specific HLA-multimer • resulted in a distinct cell cluster that did not stain with control HLA-multimers of different specificity. . • Each MLPC was considered to contain a multimer positive population.
048 and p=0.RESULTS • EBV-specific CTL responses were detected in the peripheral blood of 8/19 lung cancer patients (42%) and 5/14 (36%) aged-matched controls (p=0.713).031. respectively) that presented with an EBV-specific CTL response . • Both of these proportions were statistically significantly different than 86% (6/7) of younger healthy individuals (p=0.
only if staining with the specific HLA-multimer • resulted in a distinct cell cluster that did not stain with control HLA-multimers of different specificity.RESULTS • Each MLPC was considered to contain a multimer positive population. .
RESULTS • This indicates that all antiviral T cells had TCR with a similar avidity towards the peptide/MHC complex and no difference in the kinetics of interaction between TcR and multimer complexes could be observed .
was twice as less than that elicited by younger healthy individuals .DISCUSSIONS • This study provides direct evidence that lung cancer patients dispose an EBV-specific CTL response equivalent to that of age-matched healthy counterparts • the EBV-specific CTL response mounted by subjects of this age group. either with cancer or not.
these observations can be explained in the context of the complex process of T cell immunosenescence .DISCUSSIONS • Regarding the healthy individuals. results demonstrated an inverse correlation between age and the percentage of circulating EBV-specific CTLs. • Most likely.
cancer patients also appeared with a decreased proliferative capacity of virus specific pCTL . it is interesting that they present with the same age-related alteration of EBV-specific CTL response as their healthy counterparts • Beyond differences observed in the specific pCTL frequency related to age.DISCUSSIONS • With respect to cancer patients.
.CONCLUSIONS • this study provides evidence that lung cancer patients dispose an EBV-specific CTL response equivalent to that of age-matched healthy counterparts • study suggests that possibly the poor outcome of cancer immunotherapeutic approaches in lung cancer can be a result of the underlying effects of senescence on the immune system rather than an inefficient anti-tumor response.
EBV PROMOTES HUMAN CD8+ NKT CELL DEVELOPMENT Karen Cas .
INTRODUCTION • NKT cells • unconventional T cells • recognize CD1d-presented glycolipids .
CD8+ (SP) .or CD4.CD8.(DN) • CD4+ CD8+ (DP) • CD4+ CD8.[MICE] αβT cell development in THYMUS proceeds through 3 major stages: • CD4.
[MICE] POSITIVE SELECTION CORTICAL THYMOCYTES Semi-invariant αβTCR DP NKT precursors CD1d ligand complex THYMIC DENDRITIC CELLS NEGATIVE SELECTION .
bone marrow. gut) Final NKTdifferentiation step THYMUS . spleen.PERIPHERY (liver. lymph nodes. lung.
[MICE] • It is believed that there are no CD8+ NKT cells .
[HUMAN] • CD8 is expressed on a minor proportion of human NKT cells • CD8 marker is usually acquired after egress from the thymus .
MATERIALS AND METHODS .
TETRAMERS AND OTHER REAGENTS • Latent EBV-infected [EBV+(La)] .PATIENTS. CELLS.healthy EBV seropositive individuals • Normal control subjects (NS) – healthy seronegative individuals .
PATIENTS. CELLS. TETRAMERS AND OTHER REAGENTS Patients with EBV-associated acute infectious mononucleosis [lytic phase] ] EBV+(IMa) Diagnosed by a monospot test. and Detection of capsid-specific serum IgM Followed up at 1 year [latent phase] EBV+(IMy) .
CELLS. TETRAMERS AND OTHER REAGENTS Patients with EBVassociated Hodgkin lymphoma EBV+(HL) Diagnosed according to WHO criteria.PATIENTS. and Staged according to the Ann Arbor classification .
PATIENTS. TETRAMERS AND OTHER REAGENTS • in. CELLS. • newly diagnosed • no previous treatment • Informed consent were provided .or out-patients in different hospitals in Hubei Province in China.
CELLS. TETRAMERS AND OTHER REAGENTS Human fetal thymic cells Bone marrow (BM) cells Peripheral blood mononuclear cells (PBMC) Liver cells Voluntarily elective pregnancy terminations (<24 wk gestation. HLA typing matched HLA-A2 & HLA-DRB1) .PATIENTS.
TETRAMERS AND OTHER REAGENTS Thymic cells BM cells PBMCs ISOLATED ALIQUOTED Maintained in the vapor phase of liquid nitrogen for further use .PATIENTS. CELLS.
TETRAMERS AND OTHER REAGENTS • THYMIC DENDRITIC CELLS were separated from thymocytes by adhesion onto plastic culture dishes .PATIENTS. CELLS.
.HUMAN THYMUS/LIVER SCID CHIMERAS • 8 wk-old female SCID mice – irradiated (300cGy/mouse) prior to cell-transplantation • Human fetal thymic cells – depleted of immature and mature NKT cells • Transplanted into the thymus of anaesthetized SCID mice.
HUMAN THYMUS/LIVER SCID CHIMERAS • Human fetal liver tissue – simultaneously implanted under the mouse kidney capsule • The chimeras were then intrathymically challenged with EBV or HTLV-1 • Repeated after 6 days • Maintained for 4wks .
FETAL THYMIC ORGAN CULTURE (FTOC) • Fetal thymus dissected into pieces of ~2mm3 • 3 pcs of tissue placed into 24-well plates with culture medium containing various stimuli • (Day 7) cultured thymus fragment was dispersed into a single-cell suspension -> stained and analyzed .
FETAL THYMIC ORGAN CULTURE (FTOC) • DP thymocytes – obtained by gently grinding freshly fetal thymus lobes • resulting suspensions -> sorted using CD8 and CD4 labeling .
REAGGREGATED THYMIC ORGAN CULTURE (RTOC) • Reaggregates were formed by mixing together the desired thymic stromal cells and DP thymocytes at 1:1 ratio with other stimuli .
FLOW CYTOMETRY • α-GalCer-loaded CD1d tetramer and αβTCR • Used to define total NKT cells • In intracellular staining for detection of perforin. different cells were resuspended in cold Dulbecco’s PBS > permeabilized -> stained with mAb specific for human perforin -> analyzed .
NS negative normal control subjects 17 newly-onset acute infectious mononucleosis patients 16 newly-diagnosed EBV-associated Hodgkin Lymphoma patients EBV+(HL) EBV+(IMa) EBV+(IMy) .1. EBV-INDUCED CD8+ NKT CELLS IN VARIOUS EBV-INFECTED INDIVIDUALS 177 eligible patients and subjects EBV+(La) 128 healthy latent EBV-infected subjects 16 EBV.
EBV-INDUCED CD8+ NKT CELLS IN VARIOUS EBV-INFECTED INDIVIDUALS Figure 1.1. Frequency of total NKT cells .
1. EBV-INDUCED CD8+ NKT CELLS IN VARIOUS EBV-INFECTED INDIVIDUALS Figure 2. Frequency of co-receptor expressing NKT cells .
EBV INDUCES INTRATHYMIC CD8+ NKT CELL DEVELOPMENT Figure 3 .2. Frequencies of NKT cells in thymus and liver from hu-thy/liv-SCID chimeras challenged with EBV or HTLV-1 .
2. EBV INDUCES INTRATHYMIC CD8+ NKT CELL DEVELOPMENT
Figure 4. Frequencies of co-receptor-expressing NKT in the unchallenged (Nil) or EBV challenged (EBV) hu-thy/liver SCID chimeras
3. EBV-INDUCED CD8+ NKT CELL DEVELOPMENT DEPENDS ON THYMIC DCS
Figure 5. Frequency of co-receptor expressing NKT cells in thymus and liver
3. EBV-INDUCED CD8+ NKT CELL DEVELOPMENT DEPENDS ON THYMIC DCS
Figure 6. Frequency of coreceptor expressing NKT cells in thymus and liver
3. EBV-INDUCED CD8+ NKT CELL DEVELOPMENT DEPENDS ON THYMIC DCS Figure 7. Frequency of coreceptor expressing NKT cells in thymus and liver .
3. EBV-INDUCED CD8+ NKT CELL DEVELOPMENT DEPENDS ON THYMIC DCS Figure 8. Frequency of coreceptor expressing NKT cells in thymus and liver .
Frequency of total NKT cells in different RTOCs. . EBV-INDUCED CD8+ NKT CELL DEVELOPMENT DEPENDS ON THYMIC DCS Figure 9.3.
. Frequency of co-receptor-expressing NKT cells in different RTOCs.3. EBV-INDUCED CD8+ NKT CELL DEVELOPMENT DEPENDS ON THYMIC DCS Figure 10.
4. EBV-INDUCED CD8+ NKT CELL DEVELOPMENT IS IL-7-DEPENDENT Figure 11. Frequencies of total NKT cells and co-receptorexpressing NKT cells in different FTOCs. .
with EBV . EBV-INDUCED CD8+ NKT CELL DEVELOPMENT IS IL-7-DEPENDENT Figure 12. Development of co-receptorexpressing NKT cells in thymus or livers from hu-thy/liv SCID chimera challenged i.4.t.
5. Perforin expressions in human and chimeric CD8+ NKT cells . EBV-INDUCED CD8+ NKT CELLS PRODUCE ABUNDANT PERFORIN Figure 13.
DISCUSSION • CD8+ fraction (up to 25%) • generated in vivo after EBV-challenge • development of CD8+ NKT cells • Promoted • Thymus • DP precursor stage • thymic DCs .
DISCUSSION • Certain pathogens are important contributors to CD8lineage commitment of NKT cells • Infection by HIV and HTLV-1 results in a decrease in NKT cells • EBV-infection promotes generation of perforin-biased CD8+ NKT cells .
DISCUSSION • Protective role of CD8+ NKT against EBV-associated malignancies has been verified • Any co-receptor-expressing human NKT cells detected in the mice • developed and differentiated post-celltransplantation .
DISCUSSION • EBV-challenge chimeras – reflects viral effects on the differentiation of CD8+ NKT cells. • IL-7 • enhancer of EBV-induced development of thymic CD8+ NKT cells • in vivo. in the hu-thy/liv-SCID chimeras • in vitro in FTOCs .
CONCLUSION • Rituximab treatment eradicated CD20-expressing B cells in patients with rheumatoid arthritis infected with EBV. • Thus rheumatoid arthritis patients have better clinical responses to Rituximab when they are infected with EBV .
. • EBV-specific CTLs decrease with age only when healthy individuals of differing age groups are compared.CONCLUSION • Lung cancer patients have EBV-specific CTLs equivalent to their age-matched healthy counterparts. • Cancer does not predispose a patient to decrease their CTL response to EBV infection.
CONCLUSION • Epstein-Barr Virus is implicated in the increased number of NKT cells • Amount of increase in NKT cells is proportional to duration of infection with EBV • EBV infection promotes generation of IFN-γ.and perforinbiased CD8+ NKT .
This action might not be possible to undo. Are you sure you want to continue?
We've moved you to where you read on your other device.
Get the full title to continue reading from where you left off, or restart the preview.