Karin Strigård

Pharmacological administration of granulocyte/macrophage-colony- stimulating factor is of significant importance for the induction of a strong humoral and cellular response in patients immunized with recombinant carcinoembryonic antigen

Abstract Eighteen colorectal carcinoma patients without macroscopic disease after surgery were immunized using recombinant (r) human (h) carcinoembryonic antigen (CEA) with (n = 9) or without (n = 9) the addition of soluble granulocyte/macrophage-colony-stimulating factor (GM-CSF). The dose of rhCEA per immunization was 100 μg (n = 6), 316 μg (n = 6) or 1000 μg (n = 6). rhCEA was given s.c. on day 1 and 80 μg/day of GM-CSF s.c. on days 1–4. The schedule was repeated six times during a period of 9 months. All patients in the GM-CSF group developed a strong rhCEA-dose-dependent IgG antibody response while only one-third of the non-GM-CSF patients mounted a weak antibody response. All patients (9/9) in the GM-CSF group developed a strong rhCEA-specific proliferative T cell response as well as type I T cells (interferon γ secretion). In 45% of the patients also a weak type II T cell response (interleukin-4 secretion) was evoked. Both MHC-class-I- and -II restricted rhCEA-specific T cells were noted. A specific cellular response (proliferation and/or cytokine secretion) against native hCEA could be found in 8/9 patients in the GM-CSF group, although at a significantly lower level than against rhCEA. In the non-GM-CSF group a weak rhCEA-specific T cell response was induced. Three patients had a proliferative response, 4 patients type I T cells and 6 patients type II T cells. No signs of autoimmune reactions were noted. Local pharmacological administration of GM-CSF seemed to be a prerequisite for the induction of a strong immunity against baculovirus-produced hCEA protein. However, the cellular response against native CEA was of a significantly lower magnitude.

Durable Carcinoembryonic Antigen (CEA)-Specific Humoral and Cellular Immune Responses in Colorectal Carcinoma Patients Vaccinated with Recombinant CEA and Granulocyte/Macrophage Colony-Stimulating Factor

Purpose: Previous studies have indicated that carcinoembryonic antigen (CEA) might be a suitable immunotherapeutic target in colorectal carcinoma (CRC). The aim of the present study was to analyze the immunological and clinical effects of vaccination with CEA together with the adjuvant granulocyte/macrophage colony-stimulating factor (GM-CSF). Experimental Design: Twenty-four resected CRC patients without macroscopic disease were immunized seven times with recombinant CEA at four different dose levels over a 12-month period. Half of the patients received GM-CSF (80 μg/day for 4 consecutive days) at each immunization. Patients were monitored immunologically for 36 months and clinically for 76 months. T-cell response was evaluated by a [3H]thymidine incorporation assay, and IgG response was determined by ELISA. Results: Minor local side effects were common. All 12 patients (100%) in the GM-CSF group developed a CEA-specific T-cell as well as an IgG response. The corresponding figures in the CEA alone group were 9 of 12 (75%) and 8 of 12 (66%), respectively. GM-CSF significantly augmented the amplitude of the T-cell response and the IgG titers. No dose–response relationship was noted. The immune responses at 12 months persisted 24 months after the last vaccination. Anti-CEA IgG titers were associated with increased survival (P < 0.05), whereas standard prognostic factors had no relationship, with the exception of serum CEA value. Conclusions: Vaccination with recombinant CEA and GM-CSF appears to be a nontoxic regimen inducing potent and durable antigen-specific IgG and T-cell response. The results of this study justify more extensive trials with recombinant CEA protein for immunotherapy of CRC.

In vivo treatment of rats with monoclonal antibodies against gamma interferon: effects on experimental allergic neuritis

ABSTRACT— To elucidate the role of gamma interferon in experimental allergic neuritis (EAN) a mouse monoclonal antibody (DB‐1) directed against rat gamma interferon was used to treat rats during different phases of the development of experimental allergic neuritis (EAN). The effects of this treatment were followed by clinical evaluation, and in some instances by immunohistochemical analysis of lymphoid organs and affected nerves for presence of MHC class II antigens and various T cell subsets. DB‐1 treatment given after onset of clinical symptoms (Day 15 after immuniozation with myelin) shortened disease duration, compared with non‐treated EAN controls. Affected nerves of DB‐1 treated animals showed reduced expression of MHC class II antigens and lower numbers of T lymphocytes within the affected nerves. In contrast, when DB‐1 treatment was given on the day of immunization (Day 0), the disease duration increased, and when given before onset of the disease (Day 9) the clinical course was not significantly affected. The results support an important role for gamma interferon in the pathogenesis of EAN.

Distribution of plasma cells secreting antibodies against nervous tissue antigens during experimental allergic encephalomyelitis enumerated by a nitrocellulose immunospot assay

The B cell response to central nervous system (CNS) myelin and myelin basic protein, as well as total numbers of IgG secreting cells, was studied in acute experimental allergic encephalomyelitis using a nitrocellulose immunospot assay. The method was able to detect single plasma cells secreting antibodies. Cells secreting antibodies against myelin antigens were detected in regional lymph node cell suspension by day 5 post-immunization (p.i.). At that time no anti-myelin antibodies were detected free in serum. Later, at day 15 p.i., specific antibody secreting cells were found in bone marrow and spleen indicating a generalization of the immune response. The B cell response became partly sequestered to the target of immune attack since an increased number of IgG secreting cells was detected among mononuclear cells recovered from the CNS. Studies of cellular secretion of antibodies rather than free levels in body fluids may be a more accurate reflection of the in vivo B cell response. These findings may be generally considered in studies of B cell mediated immunity in neuroinflammatory diseases.

Pregnancy and Guillain-Barré syndrome: a nationwide register cohort study.

In this study, we determined the relationship between Guillain-Barré syndrome (GBS) and pregnancy. By taking advantage of several nationwide registers and the availability of personal identification numbers, we calculated person-years for Swedish females aged 15-49 years in the following categories: (1) neither pregnant nor postpartum; (2) pregnant; (3) in the first month postpartum, or (4) in the first 3 months postpartum during 1973-1983. For these women, we determined the corresponding exposure status of hospital-registered GBS cases. Medical records were examined for GBS cases hospitalized during the 2-week period postpartum and 1-month period after the last menstruation. Poisson regression analysis yielded age-adjusted relative risks (RRs) of 0.86 (95% CI 0.40-1.84) for pregnant women, and 1.47 (0.54-3.99) and 2.21 (0.55-8.94) for females during the 3-month and the 30-day period after delivery. The risk for GBS seems to be lower during pregnancy and increases after delivery.

Muscle strength and endurance after surgery for primary hyperparathyroidism

OBJECTIVE To evaluate the effect of surgery on muscular strength and endurance in patients with primary hyperparathyroidism (HPT). DESIGN Prospective open study. SETTING University hospital, Sweden. SUBJECTS Nine patients undergoing HPT surgery and nine matched patients undergoing thyroid resection who acted as controls. INTERVENTIONS Concentric and eccentric endurance was evaluated with a test comprising 100 repeatedly executed muscle action at 90 degrees.s-1. Blood samples obtained before and after operation were analysed for calcium, phosphate, thyroid stimulating hormone (TSH), and parathyroid hormone (PTH) concentrations. MAIN OUTCOME MEASURES Peak torque during maximum voluntary concentric and eccentric muscle actions at 90 degrees.s-1 before, three months and one year after operation. RESULTS There were no differences in concentric and eccentric peak torque before and after operation either within or between groups. Concentric and eccentric endurance were similar in the HPT group and controls before as well as after operation. The return of calcium and PTH concentrations to their reference ranges after parathyroidectomy did not correlate with changes in concentric and eccentric peak torque. CONCLUSIONS The subjective improvement in muscle endurance which is often encountered in patients with HPT after operation is not associated with an objective increase in muscle strength or endurance as measured by isokinetic muscle performance.

Validation of Biodex system 4 for measuring the strength of muscles in patients with rectus diastasis

Abstract To investigate the validity and reliability of the Biodex system 4 in the assessment of abdominal strength in patients with rectus diastasis, we studied 10 healthy volunteers and 10 patients with rectus diastasis of more than 3 cm. We assessed test-retest reliability at 30o and 60o of extension/flexion spinal angles, respectively, and isometric muscle strength with and without the abdominal girdle. Reliability was calculated using the interclass correlation coefficient (ICC). Validity was evaluated by correlation with the International Physical Activity Questionnaire (IPAQ) and a visual analogue scale (VAS) for self-assessment of abdominal muscular strength. The Kendall-Tau and Spearman tests were used. Reliability was excellent with ICC values ranging between 0.77 and 0.97. There was no significant difference in muscular strength for tests with or without a girdle except with 60o extension. The internal validity assessed as the correlation between VAS and isometric strength (p = 0.008) was excellent, whereas there was no correlation with IPAQ. The Biodex system 4 is a valuable and reliable instrument to measure abdominal wall strength in patients with rectus diastasis. The internal validity is excellent. The incongruence between abdominal muscle strength, as measured using the Biodex system 4, and IPAQ indicates that there is no relation between general physical activity (IPAQ) and muscular strength in patients with rectus diastasis.

Early complications, pain, and quality of life after reconstructive surgery for abdominal rectus muscle diastasis : a 3-month follow-up.

AIM The aim of this study was to evaluate early complications following retromuscular mesh repair with those after dual layer suture of the anterior rectus sheath in a randomised controlled clinical trial for abdominal rectus muscle diastasis (ARD). METHODS Patients with an ARD wider than 3 cm and clinical symptoms related to the ARD were included in a prospective randomised study. They were assigned to either retromuscular inset of a lightweight polypropylene mesh or to dual closure of the anterior rectus fascia using Quill self-locking technology. All patients completed a validated questionnaire for pain assessment (Ventral Hernia Pain Questionnaire, VHPQ) and for quality of life (SF36) prior to and 3 months after surgery. RESULTS The most frequently seen adverse event was minor wound infection. Of the patients, 14/57 had a superficial wound infection; five related to Quill and nine to mesh repair. No deep wound infections were reported. Patient rating for subjective muscular improvement postoperatively was better in the mesh technique group with a mean of 6.9 (range 0-10) compared to a mean of 4.8 (range 0-10) in the Quill group (p=0.01). The pre- and post-operative SF36 scores improved in both groups. CONCLUSIONS There was no significant difference between the two surgical techniques in terms of early complications and perceived pain at the 3-month follow-up. Both techniques may be considered equally reliable for ARD repair in terms of adverse outcomes during the early postoperative phase, even though patients operated with a mesh experienced better improvement in muscular strength. ClinicalTrial.gov: 2009/227-31/3/PE/96.

Functional HLA-DR T cell epitopes of CEA identified in patients with colorectal carcinoma immunized with the recombinant protein CEA

A baculovirus-produced recombinant CEA (rCEA) protein comprising the extracellular region was used for vaccination of CRC patients with or without GM-CSF as an adjuvant cytokine. Ten patients with a significant proliferative T cell response against rCEA were selected for T cell epitope mapping. Fifteen-aa-long overlapping peptides covering the entire aa sequence of the external domain of CEA were used in a proliferation assay. In six of the patients a repeatable T cell response against at least one peptide was demonstrated. For the first time, nine functional HLA-DR epitopes of CEA were defined. Two of the peptides were recognized by more than one patient, i.e., two and three patients, respectively. Those 15-mer peptides that induced a proliferative T cell response fitted to the actual HLA-DR type (SYFPEITHI). The affinity of the native peptides for the T cell receptor was in the low to intermediate range (scores 6–19). The 15-mer peptides also contained 9-mer peptide sequences that could be predicted to bind to the actual HLA-ABC genotypes (SYFPEITHI/BIMAS). Blocking experiments using monoclonal antibodies indicated that the proliferative T cell response was both MHC class I and II restricted. The defined HLA-DR T cell epitopes were spread over the entire CEA molecule, but a higher frequency was noted towards the C-terminal. Peptides with a dual specificity may form a basis for production of subunit cancer vaccines, but modifications should be done to increase the T cell affinity, thereby optimizing the antitumoral effects of the vaccine.

Imaging of parastomal hernia using three‐dimensional intrastomal ultrasonography

Parastomal hernia is common in patients with a permanent stoma. At present there is no standard method for imaging a parastomal hernia. The aim of this study was to investigate the value of three‐dimensional intrastomal ultrasonography in differentiating between a parastomal hernia and a bulge.