1. This document discusses chronic lymphocytic leukemia (CLL) and summarizes key findings about immunobiology and B-cell receptor characteristics.
2. CLL results from the selection and transformation of B cells expressing restricted B-cell receptors (BCRs) that can be polyreactive and autoreactive.
3. Distinct CLL subgroups differ in the retention or loss of polyreactivity and autoreactivity, with retention associated with worse clinical outcomes.
Kell blood group system most important blood group system following to ABO and Rh blood group system, particularly RhD as far as immunogenicity is concerned and Its clinical importance.
Linda Cendales Composite Tissue Graft Summary Banff 2013 Meeting in BrazilKim Solez ,
Composite tissue graft summary from 12th Banff Conference on Transplant Pathology from the meeting in Comandatuba-Bahia, Brazil on August 23rd, 2013 http://cybernephrology.ualberta.ca/banff/2013
Kell blood group system most important blood group system following to ABO and Rh blood group system, particularly RhD as far as immunogenicity is concerned and Its clinical importance.
Linda Cendales Composite Tissue Graft Summary Banff 2013 Meeting in BrazilKim Solez ,
Composite tissue graft summary from 12th Banff Conference on Transplant Pathology from the meeting in Comandatuba-Bahia, Brazil on August 23rd, 2013 http://cybernephrology.ualberta.ca/banff/2013
Generation of Antibody Diversity- Quick revision from Kuby through presentationSharmistaChaitali
Immunology, Kuby's fifth edition notes for strong background in the topic, General introduction, Types of Antibody and Structure, Experiments, Mechanisms
01.13.09: Chronic Myeloid Leukemia and other Myeloproliferative Neoplasms (MPNs)Open.Michigan
Slideshow is from the University of Michigan Medical
School's M2 Hematology / Oncology sequence
View additional course materials on Open.Michigan: openmi.ch/med-M2Hematology
molecular of lymphoma new.pptx MBBS PG STUDENTssuser77fe3b
Lymphoma is a cancer that affects the lymphatic system, a part of the immune system that produces white blood cells and removes excess fluids from the body. It occurs when lymphocytes, a type of white blood cell, mutate and become cancerous cells that multiply and collect in the lymph nodes.
Lymphoma is a cancer that affects the lymphatic system, a part of the immune system that produces white blood cells and removes excess fluids from the body. It occurs when lymphocytes, a type of white blood cell, mutate and become cancerous cells that multiply and collect in the lymph A cancer of the lymphatic system.
This cancer impacts the lymphatic system which is the body's disease-fighting network. It includes the lymph nodes, spleen, thymus gland and bone marrow. The main types of lymphoma are Hodgkin's lymphoma and non-Hodgkin's lymphoma.
Symptoms include enlarged lymph nodes, fatigue, significant weight loss and fever.
Treatment may involve chemotherapy, medication, radiation therapy and rarely stem-cell transplant.
nodes. A cancer of the lymphatic system.
This cancer impacts the lymphatic system which is the body's disease-fighting network. It includes the lymph nodes, spleen, thymus gland and bone marrow. The main types of lymphoma are Hodgkin's lymphoma and non-Hodgkin's lymphoma.
Symptoms include enlarged lymph nodes, fatigue, significant weight loss and fever.
Treatment may involve chemotherapy, medication, radiation therapy and rarely stem-cell transplant.
A cancer of the lymphatic system.
This cancer impacts the lymphatic system which is the body's disease-fighting network. It includes the lymph nodes, spleen, thymus gland and bone marrow. The main types of lymphoma are Hodgkin's lymphoma and non-Hodgkin's lymphoma.
Symptoms include enlarged lymph nodes, fatigue, significant weight loss and fever.
Treatment may involve chemotherapy, medication, radiation therapy and rarely stem-cell transplant.
A cancer of the lymphatic system.
This cancer impacts the lymphatic system which is the body's disease-fighting network. It includes the lymph nodes, spleen, thymus gland and bone marrow. The main types of lymphoma are Hodgkin's lymphoma and non-Hodgkin's lymphoma.
Symptoms include enlarged lymph nodes, fatigue, significant weight loss and fever.
Treatment may involve chemotherapy, medication, radiation therapy and rarely stem-cell transplant.
A cancer of the lymphatic system.
This cancer impacts the lymphatic system which is the body's disease-fighting network. It includes the lymph nodes, spleen, thymus gland and bone marrow. The main types of lymphoma are Hodgkin's lymphoma and non-Hodgkin's lymphoma.
Symptoms include enlarged lymph nodes, fatigue, significant weight loss and fever.
Treatment may involve chemotherapy, medication, radiation therapy and rarely stem-cell transplant.
A cancer of the lymphatic system.
This cancer impacts the lymphatic system which is the body's disease-fighting network. It includes the lymph nodes, spleen, thymus gland an
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
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Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Ocular injury ppt Upendra pal optometrist upums saifai etawah
Se1.2 chiorazzi pc aecc_retreat_5-5-10_for_website
1. Immunobiology of
chronic lymphocytic leukemia
Nicholas Chiorazzi
Departments of Cell Biology and of Medicine
Albert Einstein College of Medicine
and
The Feinstein Institute for Medical Research
North Shore – LIJ Health System
2. Ig V gene sequencing Autoantigen reactivity
Rajendra Damle Charles Chu
Joy Yan Rosa Catera
Bradley T. Messmer Manuela Woelfle
Emilia Albesiano Katerina Hatzi
Angelo Valetto Zev Sthoeger
Fabio Ghiotto Eric Meffre
Franco Fais Maxime Herve
Gregg Silverman
Collaborators
Kanti R. Rai
Steven L. Allen
Jonathan E. Kolitz
Matthew Kaufman
Manlio Ferrarini
3. Chronic Lymphocytic Leukemia
Most prevalent adult leukemia of the Western world
15,340 new cases and 4,500 deaths in 2007
Disease of aging individuals (>50) with incidence
increasing with each subsequent decade
Usually affects men > women; Caucasian > African > Asian
4. Clinical dilemma
Clinical courses of patients are heterogeneous and that
of an individual patient is unpredictable
Some patients follow very benign courses living
decades and dying with the disease, not from it
Other patients follow more malignant courses
living only a few years after diagnosis, despite
therapy
CLL remains an incurable disease
Therefore, “wait and watch” / “wait and worry”
approaches are taken by clinician and patient
5. Chronic Lymphocytic Leukemia
Lymphocytosis seen in blood, but most leukemic cells are
in non-vascular areas including bone marrow, lymph
nodes, and spleen
Usually detected upon routine blood workup as an
elevated white cell or lymphocyte count (~3x1010 total
in blood)
Clonal expansion of a CD5+ B lymphocyte with low surface
immunoglobulin (Ig)
Most clones express predominantly IgM isotype surface
Ig, but ~7% are predominantly IgG or IgA, though in
those cases IgM clonal relatives can be found
6. Clonal disease of B lymphocytes
CD19
CD5
CD23
Smudge cell smIg (BCR)
CLL cells
Granulocyte
7. Take home messages
1. CLL results from the non-random selection and
transformation of B lymphocytes expressing B-cell
antigen receptors (BCRs) of restricted amino acid
structure
2. These BCRs can be poly- and auto-reactive, binding
natural as well as novel autoantigens generated by
apoptosis and other catabolic processes
3. The clinically-distinct subgroups differ in the retention
or loss of poly- and auto-reactivity, with the retention of
polyreactivity being associated with worse clinical
disease
8. Take home messages
1. CLL results from the non-random selection and
transformation of B lymphocytes expressing B-cell
antigen receptors (BCRs) of restricted amino acid
structure
9. Ig molecules and their genes
NH2
VK
CK
VH
Fab CDR3 CDR2 CDR1
CH1
Fc FR4 FR3 FR2 FR1 VH D JH CH
-S - S-
Hinge CH2
Region JH D VH
Chromosome 14
VK JH CK
CH3
Chromosome 2
COOH JK VK
Ig Molecule V Region Ig Genes
10. CLL cells differ from normal CD5+ B cells
by the overuse of certain autoreactive genes
Fais et al. J Clin Invest 98: 1659, 1998
11. CLL clones differ in the degree of somatic
mutations, especially in particular IgV genes
VH Specific % Cases with
Family VH Gene Mutations
All cases - 50.7
1 - 33.3
1-69 10.0
3 - 66.7
3-07 90.0
4 - 41.2
4-34 55.0
Fais et al. J Clin Invest 98: 1659, 1998
12. Ig VH gene mutation status of CLL cells is an
important prognostic indicator of outcome
≥ 2% mutation ≥ 2% mutation
< 2% mutation < 2% mutation
Damle et al. Hamblin et al.
Blood 94: 1840, 1999 Blood 94: 1848, 1999
13. Ig molecules and their genes
NH2
VK
CK
VH
Fab CDR3 CDR2 CDR1
CH1
Fc FR4 FR3 FR2 FR1 VH D JH CH
-S - S-
Hinge CH2
Region JH D VH
Chromosome 14
VK JH CK
CH3
Chromosome 2
COOH JK VK
Ig Molecule V Region Ig Genes
14. CLL clones are culled from the normal B-cell
repertoire based on structural constraints of
the B-cell antigen receptor
15. IgV gene segment recombination
Heavy Chain Light Chain (k/λ)
VH (44) D (27) JH (6) VL (46/36) JL (5/7)
= RAG mediated
recombination
HC = VH x D x JH = 44 x 27 x 6 = 7,128 VHDJH rearrangement: ~1 : 7,000
16. ~1% of CLL patients express a BCR with a VH 1-69
gene exhibiting very similar HCDR3s often
comprised of the same VH-D-JH segments
Widhopf et al. Blood 104: 2499-2504, 2004
17. Ig V region gene segment recombination
Heavy Chain Light Chain (k/λ)
VH (44) D (27) JH (6) VL (46/36) JL (5/7)
= RAG mediated
recombination
HC = VH x D x JH = 44 x 27 x 6 = 7,128
k = Vk x Jk = 46 x 5 = 230
VHDJH / VLJL rearrangement: ~1 : 3 x 106
l = Vl x Jl = 36 x 7 = 252
20. Because of the differences that occur at the
junctions when gene segments combine, the
likelihood that the same VHDJH- VLJL
rearrangement with the same junctional
characteristic would occur in two different B
cells is even much more remote
≈ 1 / 1x108 – 1 : 1x1012
Therefore, if the gene structure of the Ig variable
region found in B-CLL cells from different
patients is very similar or identical, then this
must indicate a selective process of
leukemogenesis that targets B cells with a given
type of Ig V region.
21. Heavy chain sequence alignment
VH1-69 D3-16 JH 3
CAR YYDYVWGSYRY DAFDIWG
CLL068: CAR GG DYDYVWGSYRS N DAFDIWG
CLL258: CAR GG IYDYVWGSYRP N DAFDIWG
CLL022: CAR GG DYDYVWGSYRP N DAFDIWG
CLLSMI: CAR GG NYDYIWGSYRS N DAFDIWG
aCLA*: CAR GG NYDYIWGSYRS N DAFDIWG
Natural autoantibody
*aCLA = anti-cardiolipin ab Messmer et al. J Exp Med 2004; 200: 519-525
22. Almost 30% of patients with chronic lymphocytic leukemia
carry stereotyped receptors
Stamatopoulos et al. Blood 109:259-270, 2007
Murray et al. Blood 111:1524-1533, 2008
>35% chance of fitting into a stereotypic set if U-CLL
or if express a specific VH gene (1-69, 3-21, 4-39)
associated with poor outcome
23. Take home messages
2. These BCRs can be poly- and auto-reactive, binding
natural as well as novel autoantigens generated by
apoptosis and other catabolic processes
24. Expression of recombinant CLL mAbs
Transfection of 293T HEK cells using Lipofectamine 2000
1. Plasmid DNA 2.Lipofectamine 3. Lipofectamine 2000 4. Liposomes are added in
carrying heavy and 2000 Reagent Reagent and DNA are 293T HEK cell culture
light chain Ig gene mixed and incubated
Immuno assay for
quantification of
CLL mAb
4-5 days of culture
293T HEK cell line
Antibody purification using
Protein G beads
Wardemann et al. Science 301:1374, 2003
26. Reactivity of recombinant mAbs with viable HEp-2
cells that were permeabilized to allow Ab entry
[“Anti-cell antibodies”]
~20% of mAbs from CD5+ B cells from normal subjects
~75% of mAbs from CLL cells
~90% from unmutated CLL
~60% from mutated CLL
Reactivity mainly with cytoplasmic structures and
occasionally with nucleoli. Only one mAb from
M-CLL patient reacted with nucleus in a
homogeneous pattern
Herve et al. J Clin Invest 115:1636-1643, 2005
27. Autoreactivities of individual CLL BCRs/mAbs
1. VH1-69/D3-16/JH3 + VK3-20:
Non-muscle myosin heavy chain IIA
(Chu et al. Blood 112: 5122-5129, 2008)
2. VH4-39/D6-13/JH5 + VK1D-39/JK1
Vimentin
(Chu et al. Blood 112: 5122-5129, 2008)
3. 60% of U-CLL and 10% of M-CLL:
Apoptosis-associated autoantigens
(Catera et al. Mol Med 2008; 14: 665-674 )
29. Heavy chain sequence alignment
VH1-69 D3-16 JH 3
CAR YYDYVWGSYRY DAFDIWG
CLL068: CAR GG DYDYVWGSYRS N DAFDIWG
CLL258: CAR GG IYDYVWGSYRP N DAFDIWG
CLL022: CAR GG DYDYVWGSYRP N DAFDIWG
CLLSMI: CAR GG NYDYIWGSYRS N DAFDIWG
aCLA*: CAR GG NYDYIWGSYRS N DAFDIWG
Natural autoantibody
*aCLA = anti-cardiolipin ab Messmer et al. J Exp Med 2004; 200: 519-525
30. mAb 068 binds 225KDa molecule
C. Chu et al. Blood 112: 5122-5129, 2008
32. CLL mAb 068 co-localizes with pAbs to MYHIIA
C. Chu et al. Blood 112: 5122-5129, 2008
33. Autoreactivities of individual CLL BCRs/mAbs
1. VH1-69/D3-16/JH3 + VK3-20:
Non-muscle myosin heavy chain IIA
(Chu et al. Blood 112: 5122-5129, 2008)
2. VH4-39/D6-13/JH5 + VK1D-39/JK1
Vimentin
(Chu et al. Blood 112: 5122-5129, 2008)
3. 60% of U-CLL and 10% of M-CLL:
Apoptosis-associated autoantigens
(Catera et al. Mol Med 2008; 14: 665-674 )
34. CLL mAbs react with apoptotic (not healthy) cells
A B
3.70 28.17 16.77 13.86
Annexin V
RAMOS
67.27 0.92 69.14 0.23
CLL014 DO13
C D
52.86 18.12 55.35 16.96
Annexin V
Jurkat
28.47 0.55 27.37 0.32
CLL014 DO13
R. Catera et al. Mol Med 14: 665-674, 2008
35. Apoptotic B and T cells are a source of
autoantigens for CLL mAbs
SUMMARY of the results:
- More than 60% (18/28) of the mAbs tested reacted with
these two cell types
- 15 of the 18 reactive mAbs used an unmutated VH gene,
only 3 used a mutated VH gene
R. Catera et al. Mol Med 14: 665-674, 2008
36. Antigens bound at the surface of apoptotic cells
have translocated from intracellular compartments
Cytox Orange Annexin V CLL 114 Merge
Membrane blebs
Apoptotic body
without DNA
Apoptotic body
with DNA
R. Catera et al. Mol Med 14: 665-674, 2008
37. MYHIIA is one of the intracellular antigens
that translocates to the surface and
is bound by CLL mAbs
38. MEAC: MYHIIA exposed apoptotic cell
Late apoptotic
Live
Early apoptotic
Chu et al. Blood 2010 in press
39. CLL 068 mAb binds to MEACs
Negative Apoptotic MEACs
Chu et al. Blood 2010 in press
40. Many CLL mAbs bind MEACs
MEAC binding
Subset
Mutation
Chu et al. Blood 2010 in press
41. Take home messages
3. The clinically-distinct subgroups differ in the retention
or loss of poly- and auto-reactivity, with the retention of
polyreactivity being associated with worse clinical
disease
42. Polyreactivity is a feature primarily of unmutated CLL cells
Herve et al. J Clin Invest 115:1636-1643, 2005
43. Ig VH gene mutation status of CLL cells is an
important prognostic indicator of outcome
≥ 2% mutation ≥ 2% mutation
< 2% mutation < 2% mutation
Damle et al. Hamblin et al.
Blood 1999; 94: 1840 Blood 1999; 94: 1848
44. Binding well to MEACs correlates with poor
patient survival
Lo binding
?? Months (n = 9)
Hi binding
99 months
(n = 15)
Chu et al. Blood 2010 in press
45. In this limited series, MEAC binding correlates better
with patient survival than IGHV mutation status
Mutated
?? Months (n = 6)
Unmutated
118 months
(n = 18)
Chu et al. Blood 2010 in press
46. Many CLL mAbs bind MEACs
MEAC binding
Subset
Mutation
Chu et al. Blood 2010 in press
47. Inferences
1. MEACs may be a source of autoantigens
driving CLL disease
2. The origin of many CLL clones may be cells
that produce natural antibodies to MEACs
or other natural products of catabolism
B1-like cells, MZ B cells?
3. If MEAC binding is a “better” predictor
of patient survival than IGHV mutations
status, is it because the former implies
antigen-binding activity whereas the latter
directly measures it?
48. B-CLL Initiation
M-CLL
evolution U-CLL
hypothesis Progression
MEACs
MYHIIA+
Vimentin
Filamin B
Oxidation
Chemical
Modification
Editor's Notes
I would like to thank the organizers for giving me the opportunity to discuss our research results with you. This represents the work of not only myself but of many others including… Lu, Katerina, Rosa Steve and Kanti for clinical collaboration Nick for support, guidance and inspiration and others that I will acknowledge throughout the talk
This is a typical example of flow cytometry data showing that CLL 068 mAb reacts to MEACs. This is a negative control. This shows that CLL 068 binds to a subset of dead cells (AV-PE positive) and not live cells. This shows that CLL 068 binds only to MEACs (MYHIIA positive). And not other cells.
This is a graph that shows the binding of 26 CLL mAbs listed on the x-axis to MEACs… … with the cutoff of 1.5 for MEAC binding. 16 mAbs bind well/. This is a grouping of the CLL mAbs based on having a shared common “stereotyped” sequence, which I do not have time to describe, … but would just like to point out that the stereotyped groups bind in the same manner. This shows which CLL mAbs are mutated or unmutated, … U = is less than 2% mutation in the IGHV, which correlates with bad patient survival … M = is greater than 2% mutation in the IGHV, which correlates with good patient outcome. All the MEAC binding mAbs are UM, except for 1 (15/16), which binds like the same stereotype subgroup “1” and is a borderline “Mut”. Those mAbs that do not bind MEACs well are a mixture of Mut (6) and four UM (4) mAbs. Because survival correlates with IGHV mutation, perhaps MEAC binding correlates with survival and help distinguish the UM that may survive better.. .
Indeed binding well to MEACs correlates to poor patient survival (24 patients have survival data)… Hi binding has a median survival of 99 months (n=15), Whereas Lo binding has not reached median survival (n=9). This is statistically significant P<0.0087. This significance is better than that for UM versus Mut (P<0.06) in this patient cohort. … this is because one mutated (CLL 154) and two unmutated (CLL 376 and 412) IGHV CLL patients having survival outcomes contrary to that expected for their IGHV mutation status.
IGHV mutation status versus patient survival (24 patients have survival data)… UM IGHV has a median survival of 118 months (n=18), Whereas Mut IGHV has not reached median survival (n=6). This is not quite statistically significant P<0.06. This significance is not as good as MEAC binding. … this is because one mutated (CLL 154) and two unmutated (CLL 376 and 412) IGHV CLL patients having survival outcomes contrary to that expected for their IGHV mutation status.
This is a graph that shows the binding of 26 CLL mAbs listed on the x-axis to MEACs… … with the cutoff of 1.5 for MEAC binding. 16 mAbs bind well/. This is a grouping of the CLL mAbs based on having a shared common “stereotyped” sequence, which I do not have time to describe, … but would just like to point out that the stereotyped groups bind in the same manner. This shows which CLL mAbs are mutated or unmutated, … U = is less than 2% mutation in the IGHV, which correlates with bad patient survival … M = is greater than 2% mutation in the IGHV, which correlates with good patient outcome. All the MEAC binding mAbs are UM, except for 1 (15/16), which binds like the same stereotype subgroup “1” and is a borderline “Mut”. Those mAbs that do not bind MEACs well are a mixture of Mut (6) and four UM (4) mAbs. Because survival correlates with IGHV mutation, perhaps MEAC binding correlates with survival and help distinguish the UM that may survive better.. .
These results lead to the following inferences…
Just to end with a MODEL cell death (CLL or other) leading to MEAC formation and exposure of MYHIIA and other epitopes observed by Rosa This could be important for the initiation of CLL or the ongoing stimulation that may be required for CLL growth and development.