MONOCYTE CHEMOTACTIC PROTEIN-1

The effect of Monocyte Chemotactic Protein-1 gene polymorphisms on severe acute pancreatitis

Table of Contents

Ch # 1: Introduction3

Pathophysiology4

i) Pathogenesis4

ii) Cause of Systemic manifestations4

iii) Inflammatory Mediators5

iv) Prognosis5

v) Triggers for systemic responses5

vi) Current Therapeutic Opportunities6

Monocyte Chemotactic Protein - 1 (MCP-1)6

i) The Chemokine Superfamily9

ii) MCP-19

iii) MCP-1 Dimer Formation10

Possible Role In Acute Pancreatitis12

AIMS13

Ch # 2: Literature Review15

Rate of fibrosis progression24

DNA isolation and MCP- 1 genotyping25

Isolation and stimulation of activated HSC25

MCP- 1 and IL-8 secretion of activated HSC26

Isolation of hepatic RNA and quantification of MCP- 1 expression27

Gel mobility-shift assays27

Ch # 3: Methodology29

Patients And Methods29

i)Inclusion Criteria29

ii)Exclusion Criteria29

iii)Inclusion Procedures29

iv)Facilities, Personnel and Practical Issues30

v)Data Collection:30

vi)Measurement of serum Monocyte Chemo attractant Protein-1 levels31

vii)Measurement of urinary Monocyte Chemo attractant Protein-131

vii)Comparison of Monocyte Chemotactic potency between wild and -2518 variant MCP-1 types.31

viii)rt-PCR Analysis of MCP-132

xi)Statistical Analysis, Sample Size and Power32

Ch # 4: Data Analysis34

Statistical Analysis34

Demography of Data34

Cross Tabulations & Chi square36

Oneway ANOVA49

Paired sample T test50

Unpaired T test51

MCP- 1 concentrations in relation to age and lifestyle variables51

Frequency of the -2518 MCP- 1 polymorphism in HCV patients with different rates of progression of fibrosis53

Frequency of the -2518 MCP- 1 polymorphism in HCV patients with different degree of inflammation54

Phenotypic manifestation of the functional -2518 MCP- 1 polymorphism in the liver54

MCP- 1 expression in activated HSC isolated from individuals with different MCP- 1 genotypes56

MCP- 1 concentrations in relation to biochemical phenotypes57

MCP- 1 concentrations in relation to genotypes58

Ch #5: Discussion & Conclusion67

Conclusion82

References84

Ch # 1: Introduction

Monocyte chemotactic protein-1 (MCP- 1) , a specific factor that chemoattracts and activates monocytes and macrophages, is a 76-amino acid protein that is produced by many types of cells including monocytes, lymphocytes, fibroblasts, and smooth muscle cells . Activated monocytes/ macrophages secrete many angiogenic and other growth factors, which may promote the growth of endometrial explants and the expressions of numerous proinflammatory molecules. Moreover, MCP- 1 has been one of the most commonly investigated chemokines during recent years, because in women with endometriosis, the monocyte/ macrophage system is considered to play a central role in the maintenance of humoral and cell-mediated immunity, and it has been reported that the activity of MCP- 1 is elevated in endometriosis patients . Recently, several investigators reported a relationship between serum or peritoneal fluid , and MCP- 1 levels and endometriosis, but published results are conflicting.

Acute Pancreatitis is an acute inflammatory disease which has an incidence of 30-40 per 100,000 in the UK.

In the United Kingdom, mortality for acute pancreatitis has remained static at 8-10 % for the last 40 years 1. There is a spectrum of severity ranging from mild and self-limiting to severe in up to 30 % of cases.

Most patients demonstrate systemic manifestations which reflect the systemic inflammatory response. These include fever, tachycardia, tachypnoea, hypotension and oliguria. Notably in severe attacks persistence of MODS greater than > 48 hours are associated with an especially high mortality 6, 7.

Severe acute pancreatitis is associated with a marked systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome (MODS), progression to multiple organ system failure (MOSF) and ...