Detection Of Anti-carbonic Anhydrase Antibody, CD44 And CRP In Nephrotic Syndrome, Chronic Glomerulonephritis And 2-diabetic Nephropathy

Kidney diseases are uropoiesis system diseases that harm to people'shealth serious. They are not only hard to cure, but also occur repeatedly.When developing to end stage renal diseases they have a high mortality.With the development of basic subjects such as molecule cell biology andmolecule immunology, people have taken a wide and deep research onthe pathogenesis of kidney diseases, humoral immunity and cellimmunity are accepted gradually by people, but the real pathogenesis isnot elucidated fully.carbonic anhydrase(CA) is a kind of important metalloenzyme thatis widely distributed in the body and is involved in many physiologicalprocesses for example regulation of pH, transport of the CO₂ and HCO₃^-,secretion of H⁺ and so on, especially has a correlation with the normalfunction of the kidney. CD44 has played a wide role in hemopoiesis,homing and immigration of lymphocyte, stimulus and activation oflymphocyte, activation and induction of the death of cell. At the sametime, CD44 participates in the development of many kidney diseases as aadhesion molecule and correlates with infiltration of inflammation,proliferation of mesenterium cell, injury of renal tubule interstitial tightly.CRP is a kind of acute phase protein and it can reflect the level ofinflammation of the body.This experiment begins with two sides mentioned above, in order todiscuss the relationship between the pathogenesis of kidney diseases andanti-carbonic anhydraseâ…¡(anti-CAâ…¡) antibody, CD44, hope that it willhelp to get deeper understanding of the development of kidney diseasesand supply dependable proof to elucidate the pathogenesis of kidneydiseases.Part One Detection of CD44 in flow cytometry in patients with withNS, CGN and 2-DNObjiective: To discuss the level and significance of the CD44 inplasma with NS, CGN and 2-DN, evaluate the role of the CD44 played in the development of this three kinds of kidney diseases and discuss thepathogenesis in the molecule level of this three kidney diseases.Method: 76 patients were recruited for the study as control,including NS group(25), CGN group (31) and 2-DN group(20). Normalcontrol 1(NC1) was to match NS and CGN groups, Normal control 2(NC2) to match 2-DN group. the percentage of peripheral mononuclearcells which express CD44 in plasma was examined by the molecule probein flow cytometry. Serum albumin, cholesterol, creatinine and blood ureanitrogen were examined by the automatic biochemistry analyzer.Result: 1. Compared with that in the NC1 group(33.28±3.42%),there was significantly higher ratio of the peripheral mononuclear cellswhich express CD44 in patients with NS (46.47±5.54%)(p＜0.05) andCGN (58.60±4.14%)(p＜0.05). Compared with the NC2 group (23.90±3.42%), there was significantly higher ratio of that in patients with 2-DN(55.45±5.04%)(p＜0.01) 2. Relative analysis demonstrated that serumCD44 in NS group was positively associated with serum cholesterol andnegatively correlated with serum albumin, while serum CD44 in CGNand 2-DN groups was positively associated with creatinine and bloodurea nitrogen.Conclusion: the percentage of peripheral mononuclear cell whichexpress CD44 in plasma elevates in NS, CGN and 2-DN and relativeanalysis demonstrated that serum CD44 in NS group was positivelyassociated with serum cholesterol and negatively correlated with serumalbumin, while serum CD44 in CGN and 2-DN groups was positivelyassociated with creatinine and blood urea nitrogen. All the results suggestCD44 plays an important part in the development of these renal diseasesin cell immunity and will be helpful indexes in diagnosis of these kidneydiseases. Part two The building and application of ELISA for anti-CAâ…¡inplasma and the detection of C-reactive protein with NS, CGN and2-DNObjective: To discuss the level and significance of the titer ofanti-CAâ…¡antibody in plasma with NS, CGN and 2-DN, evaluate the roleplayed in the development of this three kinds of kidney diseases anddiscuss the pathogenesis in the humoral immunity of kidney diseases.Method: To build a method of indirect ELISA for detectinganti-CAâ…¡antibody and detect the concentration of CRP in plasma withimmunonephelometric methods.Results: 1. The optimum measurement conditions for ELISA is thatthe dilution of the subjects' plasma is 1:100 and the dilution of the labeledsecondary antibody is 1:200. 2. Compared with the NC1 group (3165.97±511.73), there was significantly higher plasma anti-CAâ…¡antibody titerin patients with the CGN group(4777.65±539.19)(p＜0.05). Althoughantibody titer mean in patients with NS group(1907.05±390.64)waslower than NC1 group, there was no significant difference (p＞0.05).Compared with NC2 Group(1037.17±119.42), there is significantlyhigher plasma anti-CAâ…¡antibody titer in patients with 2-DN(6875.54±1203.45)(p＜0.01). The plasma CRP in patients with NS (3.16±0.51mg/L)and CGN(2.50±0.38 mg/L) compared with NC1 group (1.85±0.46mg/L)was no significant difference (p＞0.05). Compared with theNC2 group(3.08±0.82mg/L), there was significantly higher CRP inpatients with 2-DN(7.02±0.92 mg/L)(p＜0.05).Conclusion: 1. It is possible to build a method of indirect ELISAfor detecting anti-CAâ…¡antibody. 2. These results revealed that CAâ…¡maybe a target antigen inducing immunologic reaction in CGN and 2-DN.Here, we demonstrated anti-CAâ…¡antibody and CRP play an importantrole in the development of these renal diseases in humoral immunity.