INTRODUCTIONWith advancement in the understanding of the pathogenesis underlying diabetes mellitus (DM), the boundary between type 1 and type 2 DM (T1DM and T2DM) does not seem to be as clear cut as previously thought. This study was designed to test the possibility of overlap between the spectra of immune-mediated DM and insulin resistance.

METHODSTo test for the possibility of overlap, we looked for autoantibodies typical of T1DM in patients with classical T2DM, and insulin resistance in patients with T1DM. Autoantibodies against islet cell antigen, glutamic acid decarboxylase-65 and insulinoma-associated antigen-2 were tested in 82 patients with T2DM and 27 patients with T1DM. The patients had been diagnosed on clinical criteria using standard laboratory techniques. Clinical parameters of diagnostic importance were noted, and homeostatic model assessment of insulin resistance (HOMA-IR) was calculated using fasting insulin and fasting blood glucose ratio.

RESULTSAutoantibodies against one or more beta cell antigens were detected in 12.19% of patients clinically diagnosed to have T2DM, and insulin resistance (HOMA-IR > 2.5) was diagnosed in 37.03% of patients with T1DM. It was not possible to identify any combination of clinical or biochemical markers that could predict autoantibody positivity in T2DM patients. T1DM patients with insulin resistance had a significantly higher body mass index than their insulin-sensitive counterparts (p = 0.02).

CONCLUSIONAutoantibodies against beta cell antigens are detectable in insulin-resistant T2DM patients, and insulin resistance may be present in relatively overweight T1DM patients. No differentiating clinical features that might predict autoantibody positivity in T2DM patients were found.

Adolescent ; Adult ; Autoantibodies ; blood ; immunology ; Biomarkers ; blood ; Blood Glucose ; analysis ; Body Mass Index ; Cross-Sectional Studies ; Diabetes Mellitus, Type 1 ; classification ; immunology ; Diabetes Mellitus, Type 2 ; classification ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Insulin Resistance ; immunology ; Male

OBJECTIVETo investigate the relation between insulin resistance and glutamic acid decarboxylase antibody (GAD-Ab) titers in latent autoimmune diabetes in adults (LADA).

METHODSThe patients with phenotypic type 2 diabetes were screened for GAD-Ab positivity, and the 141 positive patients were divided into two subgroups according to the GAD-Ab titer, namely the high-titer group (LADA-1 subtype) and low-titer group (LADA-2 subgroup). The clinical features and insulin resistance were compared between the two groups. Insulin resistance was calculated by HOMA 2 software, and GAD-Ab and C peptide were determined with radioligand and radioimmune assay, respectively.

RESULTSCompared with low-titer LADA patients, the patients with high titers had younger age of onset, lower BMI, higher HbA1c level, and worse fasting and postprandial C peptide levels. The insulin resistance index by HOMA 2 was significantly lower in LADA-1 group than in LADA-2 group (1.6-/+1.1 vs 2.1-/+1.1, P=0.001). The HOMA2-IR index showed a negative correlation to GAD-Ab titer.

CONCLUSIONThe degree of insulin resistance is correlated to GAD-Ab titers in LADA, and low titer patients have higher insulin resistance level.

Adult ; Aged ; Autoantibodies ; blood ; Autoimmune Diseases ; diagnosis ; immunology ; Diabetes Mellitus, Type 2 ; diagnosis ; immunology ; Female ; Glutamate Decarboxylase ; immunology ; Humans ; Insulin Resistance ; Islets of Langerhans ; immunology ; physiology ; Male ; Middle Aged

OBJECTIVETo investigate the diagnostic value of serum islet autoantibody-glutamic acid decarboxylase antibody (GADA) and islet cell antibody (ICA) in patients with hepatogenic diabetes.

METHODSSerum GADA and ICA were measured with enzyme-linked immunosorbent assay (ELISA) in 217 patients with chronic hepatitis B (CH) or liver cirrhosis (LC). The positivity rate of GADA and ICA in different phases of CH and LC and their relations with diabetes mellitus were analyzed.

RESULTSThe positivity rate of the islet autoantibody in the circulation was 72% in CH and LC patients with diabetes mellitus and 30% in patients with normal glucose level, showing significant difference between the two patient groups (Chi2=36.620, P=0.000). CH patients with diabetes had much higher positivity rate for the antibody [52% than type 2 diabetic patients with liver dysfunction [8%, P<0.05]. The positivity rate was also much higher in CH and LC patients with lowered C peptide level [70%] than in those with normal C peptide level [40%, P<0.005].

CONCLUSIONBoth GADA and ICA have important value in the diagnosis of hepatogenic diabetes and may serve as indexed in laboratory test for distinguishing hepatogenic diabetes from type 2 diabetes.

Adult ; Autoantibodies ; blood ; Diabetes Mellitus, Type 1 ; complications ; diagnosis ; immunology ; Diabetes Mellitus, Type 2 ; complications ; diagnosis ; immunology ; Diagnosis, Differential ; Female ; Glutamate Decarboxylase ; immunology ; Hepatitis B, Chronic ; complications ; Humans ; Islets of Langerhans ; immunology ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Predictive Value of Tests

OBJECTIVETo explore the role of the autoantibodies against M(2)-muscarinic receptor (M(2)-receptor), beta(1)-adrenergic receptor (beta(1)-receptor) in the development of diabetic with refractory hypertension.

METHODSSerum autoantibodies against M(2) and beta(1) were detected by ELISA using synthesized epitopes of the second extracellular loop of M(2) receptor (169 - 193) and beta(1) receptor (197 - 222) in healthy controls (n = 40), diabetic patients (n = 62), diabetic patients with non-refractory hypertension (n = 55) and diabetic patients with refractory hypertension (n = 81).

RESULTSThe positive rates of the autoantibodies against M(2) receptor and beta(1) receptor were similar among healthy controls (15.0% and 17. 5%), diabetes mellitus patients (17.7% and 14.5%) and diabetic patients with non-refractory hypertension (16.4% and 12.7%) but are significantly higher in diabetic patients with refractory hypertension (64.2% and 55.6%, P < 0.01 vs. other 3 groups).

CONCLUSIONThis finding suggests that autoimmune mechanisms might play a role in the pathogenesis of diabetic patients with refractory hypertension.

Adult ; Autoantibodies ; blood ; Diabetes Mellitus, Type 2 ; blood ; complications ; Female ; Humans ; Hypertension ; blood ; complications ; Male ; Middle Aged ; Receptor, Muscarinic M2 ; immunology ; Receptors, Adrenergic, beta-1 ; immunology

OBJECTIVETo explore the relationship between the T cell subsets and glucose level and first-phase insulin secretion function in patients with type 2 diabetes mellitus (T2DM).

METHODSWe determined the oral glucose tolerance (OGTT), insulin release test(IRT), body mass index(BMI), glycohemoglobin A1c (HbA1c), T lymphocyte subsets (CD4(+),CD8(+)), and activity of natural kill(NK) cell and ⊿I(30)/⊿G(30) in 78 newly diagnosed T2DM patients, 60 impaired glucose tolerance (IGT) patients, and 60 normal controls.

RESULTSDM and IGT patients had significantly lower levels of CD4(+), CD4(+)/CD8(+)ratio, activity of NK cell, and ⊿I(30)/⊿G(30) and significantly higher levels of HbA1c and CD8(+)compared with normal controls(all P<0.05). Patients in DM group had significantly lower level of CD4(+),⊿I(30)/⊿G(30) and significantly higher levels of FBG and HbA1c compared with IGT group. There was no significant difference in terms of CD8(+), CD4(+)/CD8(+)ratio, and activity of NK cell between IGT and DM groups, whereas CD4(+) T cells were negatively correlated with FBG and HbA1c and positively with ⊿I(30)/⊿G(30) . Multiple regression stepwise analysis showed that CD4(+) was independently associated with HbA1c and ⊿I(30)/⊿G(30).

CONCLUSIONT2DM patients tends to have disorders in cellular immunity, which is correlated with blood glucose level and the insulin secretion function.

Adult ; Aged ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; immunology ; Female ; Humans ; Immunity, Cellular ; Insulin ; blood ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo investigate the effect of laparoscopic Roux-en-Y gastric bypass on the short-term immune function of type 2 diabetic patients and to explore the correlation between the immune regulatory effect and blood glucose control.

METHODSClinical data of 28 patients with type 2 diabetes mellitus who underwent laparoscopic Roux-en-Y gastric bypass surgery in our hospital during January 2014 to January 2015 were retrospectively collected. Fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), immunoglobulin (IgG, IgA, IgM), T lymphocytes CD3(+), killer T cells CD3(+)CD8(+), helper T cells CD3(+)CD4(+), natural killer(NK) cell subsets CD16(+),CD56(+) and B cell subsets CD19(+),CD45(+) levels were detected and compared between before and after surgery. Correlation of immunoglobulin and immune cell subsets with the level of FBG and HbA1c were examined.

RESULTSAfter operation, FBG and HbA1c decreased significantly from (5.78±1.15) mmol/L to (14.21±1.89) mmol/L, and (9.96±0.97)% to (6.87±0.69)%, respectively (all P<0.05), and immunoglobulin (IgG, IgA, IgM) increased significantly [IgG: from(9.41±1.23) g/L to (12.74±1.61) g/L, IgM: from (1.71±0.22) g/L to (2.43±0.39) g/L, IgA: from (1.25±0.26) g/L to (1.97±0.23) g/L, all P<0.05]. Besides, T lymphocytes CD3(+), killer T cells CD3(+)CD8(+) and B cell subsets CD19(+)CD45(+) elevated significantly as well [T lymphocytes CD3(+): from (55.3±3.8)% to (67.6±4.6)%, killer T cells CD3(+)CD8(+): from (15.6±5.3)% to (28.7±4.2)%, B cell subsets CD19(+)CD45(+): from (8.4±3.7)% to (18.1±4.1)%, respectively, all P <0.05]. There was no significant difference in the expression of helper T cells CD3(+)CD4(+) and natural killer cell subsets CD16(+)CD56(+) between pre-operation and post-operation(all P>0.05). Levels of immunoglobulin (IgG, IgA, IgM), CD4(+)/CD8(+) ratio and B cell subsets were negatively correlated with HbA1c and FBG levels (IgG with FBG: r=-0.865, IgA with FBG: r=-0.887, IgM with FBG: r=-0.902, CD4(+)/CD8(+) with FBG: r=-0.956, CD19(+)CD45(+) with FBG: r=-0.834; IgG with HbA1C: r=-0.859, IgA with HbA1C: r=-0.957, IgM with HbA1C: r=-0.843, CD4(+)/CD8(+) with HbA1C: r=-0.912, CD19(+)CD45(+) and HbA1C: r=-0.885, all P<0.05), but the proportion of NK cells was not significantly correlated with HbA1c and FBG (P>0.05).

CONCLUSIONSLaparoscopic Roux-en-Y gastric bypass has good effect on the immune function of patients with type 2 diabetes mellitus. Improvement of immune function is correlated with the control of FBG and HbA1c.

Blood Glucose ; analysis ; Diabetes Mellitus, Type 2 ; immunology ; surgery ; Gastric Bypass ; Glycated Hemoglobin A ; analysis ; Humans ; Immunoglobulins ; blood ; Laparoscopy ; Lymphocyte Subsets ; immunology ; Postoperative Period ; Retrospective Studies

OBJECTIVETo further study the role of human cytomegalovirus (HCMV) infection in the pathogenesis of the type 2 diabetes mellitus.

METHODSHCMV DNA levels in sera from 29 type 2 diabetic patients and 23 controls were measured by quantitative polymerase chain reaction (PCR), and comparative analyses was made between HCMV DNA and T cell subsets, blood glucose (BG), insulin (Ins) and C peptide (C-P).

RESULTSThe levels of HCMV DNA were (1.81+/-1.67) x 10(8) copies/ml for type 2 diabetic patients, a level significantly higher than that (5.50+/-4.30) x 10(7) copies/ml of controls. The percentage of CD8 for type 2 diabetic patients with positive HCMV DNA was 29.53%+/-2.00%, being much higher than that for controls (27.13%+/-4.12%), while the ratio of CD4/CD8 1.24+/-0.05 was significantly lower than that 1.41+/-0.10 of controls. Fasting C-P of type 2 diabetic patients with positive HCMV DNA was far lower than that of those with negative HCMV DNA, but the differences of BG and Ins between the two groups were not significant.

CONCLUSIONActive HCMV infection of type 2 diabetic patients may suppress cellular immunity and its influence on the pathogenesis of the type 2 diabetes mellitus should be further studied.

Adult ; Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; complications ; immunology ; DNA, Viral ; blood ; Diabetes Mellitus, Type 2 ; immunology ; virology ; Female ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; T-Lymphocyte Subsets

BACKGROUND: Insulin assays are affected by varying degrees of interference from anti-insulin antibodies (IAs) and by cross-reactivity with recombinant insulin analogues. We evaluated the usefulness of the E170 insulin assay by assessing IA effects and cross-reactivity with 2 analogues. METHODS: Sera were obtained from 59 type 2 diabetes patients receiving continuous subcutaneous insulin infusion and 18 healthy controls. Insulin levels were determined using an E170 analyzer. To investigate the effects of IAs, we performed IA radioimmunoassays, and analyzed the differences between directly measured insulin (direct insulin) and polyethylene glycol (PEG)-treated insulins (free, IA-unbound; total, IA-bound and unbound insulin). We performed in-vitro cross-reactivity tests with insulin aspart and insulin glulisine. RESULTS: In IA-positive patients, E170 free insulin levels measured using the E170 analyzer were significantly lower than the direct insulin levels. The mean value of the direct/free insulin ratio and IA-bound insulin, which were calculated as the difference between total and free insulin, increased significantly as endogenous IA levels increased. The E170 insulin assay showed low cross-reactivities with both analogues (< 0.7%). CONCLUSIONS: IAs interfered with E170 insulin assay, and the extent of interference correlated with the IA levels, which may be attributable to the increase in IA-bound insulin, and not to an error in the assay. The E170 insulin assay may measure only endogenous insulin since cross-reactivity is low. Our results suggest that the measurement of free insulin after PEG pre-treatment could be useful for beta cell function assessment in diabetic patients undergoing insulin therapy.
Adult ; Aged ; Aged, 80 and over ; Cross Reactions ; Diabetes Mellitus, Type 2/blood/immunology ; Female ; Humans ; Infusions, Subcutaneous ; Insulin/analogs & derivatives/*blood/chemistry/immunology ; Insulin Antibodies/*blood ; Male ; Middle Aged ; Polyethylene Glycols/chemistry ; Radioimmunoassay/instrumentation/*methods ; Recombinant Proteins/analysis/immunology/metabolism

OBJECTIVE: To determine the correlation of serum soluble CD14 (sCD14) level with the injury of vascular endothelial cells and chronic low grade inflammation in newly diagnosed Type 2 diabetes (T2DM). METHODS: ELISA was used to examine serum sCD14 and serum soluble E-selectin (sE-selectin) level, while immunoturbidimetric assay was used to detect serum high sensitivity C reactive protein (hsCRP). RESULTS: The levels of serum sCD14, sE-selectin, and hsCRP in newly diagnosed T2DM group were higher than those in the euglycemic group [sCD14: (300.7+/-136.6) ng/mL vs. (273.3+/-86.0) ng/mL); sE-selectin: (21.3+/-7.7) ng/mL vs. (32.9+/-11.4) ng/mL; hsCRP: (1.45+/-1.21) mg/L vs. (2.37+/-1.45)mg/L], and there was a significant difference in the latter two parameters between the 2 groups(P<0.01). In the patients with newly diagnosed T2DM, after matching blood pressure, blood sugar, and blood lipid, the levels of serum sCD14, sE-selectin, and hsCRP in the obese group were higher than those in the non-obese group. There was no significant difference in the former 2 parameters between the 2 groups. The serum sE-selectin was correlated with fasting blood sugar (r=0.369, P<0.001), 2-hour postprandial blood sugar (r=0.421, P<0.001), glycosylated hemoglobin (r=0.291, P=0.005), sCD14(r=0.312, P=0.002), and homeostasis model assessment-insulin resistance(r=0.247, P=0.018) in the newly diagnosed T2DM group. Stepwise regression ana-lysis showed that the serum sCD14 was one of the chief influencing factors on serum sE-selectin. CONCLUSION Serum sCD14 levels tend to increase in newly diagnosed T2DM patients, especially in the obese diabetic patients, which is one of the chief influencing factors to induce the injury of vascular endothelial cells. The innate immunity mediated by Toll-like receptor 4 may take part in the injury of vascular endothelial cells in newly diagnosed T2DM patients.
Aged ; C-Reactive Protein ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; immunology ; E-Selectin ; blood ; Female ; Humans ; Lipopolysaccharide Receptors ; analysis ; blood ; Male ; Middle Aged ; Regression Analysis ; Solubility ; Toll-Like Receptor 4 ; immunology

OBJECTIVETo investigate the difference between serum true insulin (TI) and immunoreactive insulin (IRI) in evaluating islet beta-cell function and insulin resistance.

METHODSThe arginine stimulation test was performed in 141 individuals, including 35 with normal glucose tolerance (NGT) and 106 with type 2 diabetes (T2DM). Plasma glucose (PG), TI, IRI and proinsulin (PI) levels were measured; the incremental value of TI/PG, (TI+PI)/PG and IRI/PG (delta TI/PG, delta(TI+PI)/PG and deltaIRI/PG) and the area under curve of TI/PG, (TI+PI)/PG and IRI/PG (AUC [TI/PG], AUC[(TI+PI)/PG] and AUC [IRI/PG]) after arginine stimulation were calculated to evaluate beta-cell function.

RESULTThere were positive correlations of delta TI/PG with delta (TI+PI)/PG and delta IRI/PG in both NGT and T2DM patients (r=0.68 - 0.99, P<0.01). The similar correlations of AUC [TI/PG] with AUC [(TI+PI)/PG] and AUC [IRI/PG] were also shown (r=0.62 - 0.99, P<0.01). delta TI/PG was correlated with AUC [TI/PG] in two groups (NGT r=0.96, T2DM r=0.82, P<0.01). HOMA-IRTI, HOMA-IR(TI+PI) and HOMA-IRIRI in T2DM were higher than those in NGT (P<0.01). After arginine stimulation T2DM subjects mainly presented insulin resistance and decreased insulin secretion.

CONCLUSIONThe determination of TI may be more accurate than IRI in evaluating beta-cell function and insulin resistance.

Adult ; Aged ; Arginine ; pharmacology ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; Female ; Glucose Tolerance Test ; Humans ; Insulin ; blood ; immunology ; Insulin Resistance ; Insulin-Secreting Cells ; physiology ; Male ; Middle Aged ; Proinsulin ; blood