Impaired specific antibody response and increased B-cell population in transient hypogammaglobulinemia of infancy.

BACKGROUND: Transient hypogammaglobulinemia of infancy (THI) is a heterogeneous disorder with poorly understood pathophysiology. OBJECTIVES: To better characterize THI and improve understanding of its pathophysiology. METHODS: Twenty-four children with hypogammaglobulinemia defined by an IgG level less than 2 SDs below the mean on 2 occasions, who did not have other immunologic diagnoses, were followed and retrospectively reviewed. RESULTS: The average z-score for IgG level at presentation was -2.4 (mean age, 12 months; median age, 8 months), with a mean level of 254 mg/dL. Thirteen of 24 patients had IgA levels less than 2 SDs below the mean, 5 had IgM levels less than 2 SDs below the mean, and 7 of 23 had elevated IgE levels. Eighteen were followed up until their IgG levels normalized (mean age, 27 months; median age, 23 months), with 12 of 18 normalizing by 24 months and the remainder by 59 months. There was a significant association between presenting IgG z-score and duration of disease (P = .05). Five of the 18 patients had absolute CD19+ B-cell counts greater than the 95% percentile for age (P < .001), and the mean percentage and absolute CD19+ B-cell count across all patients were greater than those of the age-matched controls (P = .02). Most patients had nonprotective titers to Haemophilus influenzae type b vaccine, and one third had nonprotective titers to tetanus vaccine. Twenty patients carried at least one atopic diagnosis, and 13 of those had recurrent wheezing. CONCLUSIONS: THI is associated with a number of immunologic abnormalities beyond just hypogammaglobulinemia. These abnormalities include impaired specific antibody responses and increased proportions of CD19+ B cells and may be suggestive of particular immunologic mechanisms that result in hypogammaglobulinemia.     

B lymphocyte subsets and outcomes in patients with an initial diagnosis of transient hypogammaglobulinemia of infancy

Purpose

Transient hypogammaglobulinemia of infancy (THI ) is a common immunodeficiency, but definitive diagnosis can only be made retrospectively. While the pathogenesis is still unknown, abnormalities have been reported in the B cell compartment. In this study, we analysed the B cell subsets of patients with an initial THI diagnosis (n = 20) and compared them with those of healthy age‐matched Turkish children (n = 72).

Methods

Flow cytometric analyses of the B subsets were performed by staining with anti‐CD 27‐PE , anti‐CD 19‐PerCP , anti‐IgD‐FITC and anti‐IgM‐APC antibodies.

Results

During a median follow‐up of 6.6 years, 13 patients whose IgG levels had normalized before they reached four years of age were diagnosed with definitive THI . The memory subsets of these patients were lower but not statistically different from the healthy controls (HC ). The remaining seven patients had prolonged hypogammaglobulinemia after the age of four and had significantly lower memory B cell subsets compared to the HC . On follow‐up, these patients had not experienced recurrent infections or autoimmunity. Re‐evaluation of patients’ B cell subsets six years later showed that the memory B cell ratios had increased to levels comparable to HC , despite the patients still having mildly low IgG levels.

Conclusion

Patients with prolonged hypogammaglobulinemia had lower levels of memory B cells despite having a similar clinical course to patients who had been diagnosed with definitive THI . This subgroup of putative THI patients poses a diagnostic and classification dilemma. Our results suggested that these patients’ memory B cells and IgG levels may recover over time.

     

Transglutaminase as a marker for subsets of murine macrophages

Transglutaminase was detected either at the single cell level by fluorescent staining with dansylcadaverine or in cell homogenates by incorporation of [¹⁴C] putrescine into α-casein. In the mouse it was found that erythrocytes, granulocytes, thymocytes or lymphocytes with or without concanavalin A stimulation were negative in the fluorescence test. Normal peritoneal washout macrophages and peritoneal exudate cells stained positive to varying degrees (induced with mineral oil 64%, with thioglycollate 50%, with proteose peptone 22%, normal washout 1%). Macrophages from bone marrow liquid cultures were 20% positive at day 3 and 100% at day 17. Promonocytes and monocytes were negative. Positively stained cells also phagocytosed opsonized sheep erythrocytes. The degree of staining varied considerably in the macrophagelike cell lines IC21 (100%), J774.2 (75%), P388-D₁ (50%). This result and those from autoradiography studies indicate that expression of transglutaminase is not associated with the S-phase of the cell cycle. The fluorescence test correlates quantitatively with the [¹⁴C] putrescine incorporation test. The enzyme is Ca²⁺-dependent and appears neither to be on the outer cell surface nor being released into the culture medium. Circumstantial evidence indicates that it is also not compartmentalized in cytoplasmic vesicles. While the induction and modulation of enzyme expression is still under study, it is concluded that transglutaminase is a new marker for macrophages of a certain differentiation or activation state.     

Memory and naïve B-cell subsets in patients with multiple sclerosis

Memory and naïve B cells are considered to play distinct roles in immune regulation. However, the roles of memory and naïve B-cell subsets in multiple sclerosis (MS) have not yet been elucidated. In this study, we examined whether memory and naïve B-cell subsets differ between patients with MS and healthy subjects and whether interferon beta (IFNβ)-1b can affect these subsets in patients with MS. We also studied these subsets in relapsing and remitting stages of MS. Subjects included 31 patients with relapsing–remitting MS in the remitting stage, of which 15 were treated with IFNβ-1b and 16 were not treated, and 22 healthy control subjects. For 11 of the 16 untreated patients, blood samples were also obtained in the relapsing stage. Expression of CD5, CD80, CD86, CCR5, CXCR3, CD11a, and CD49d in memory and naïve B cells in blood samples was examined by flow cytometry. The percentages of CD86⁺ cells and CCR5⁺ cells in the naïve B-cell subset were significantly higher in untreated patients than in control subjects or IFNβ-treated patients. In patients with MS, the percentages of CD86⁺ cells and CCR5⁺ cells in the naïve B-cell subset and the percentage of CD5⁺ cells in the memory B-cell subset were significantly greater in the remitting stage than in the relapsing stage. These results indicate that memory and naïve B-cell subsets, especially CD86⁺ naïve B cells, CCR5⁺ naïve B cells, and CD5⁺ memory B cells, might be useful in the study of the pathogenesis of and therapy for MS.     

C-myc as a predictive marker for chemotherapy in metastatic breast cancer

C-myc is considered to have an important role in cancerogenesis and tumor progression. The aim of this study was to evaluate a possible significance of c-myc amplification as a clinically useful prognostic/predictive parameter in metastatic breast cancer (MBC). Eighty-seven MBC patients with known clinicopathological parameters were included in the study, at the time of diagnosis of metastatic disease. In metastatic setting, 52% of patients received CMF, 34% received FAC, and 32% received hormonal therapy (tamoxifen). C-myc amplification was analyzed by chromogenic in situ hybridization, according to the manufacturer's instructions. C-myc amplification was detected in 26% cases and showed a strong correlation with ER status, stage of disease (initial) and existence of distance metastasis. There was no statistically significant difference in MBC (post-relapse) survival between c-myc-nonamplified and c-myc-amplified subgroups regardless of or regarding the treatment. However, correlation was found between c-myc status and individual patient's outcomes. Patients with c-myc amplification treated with chemotherapy (CMF and FAC) had clinical benefit (complete remission, partial remission or stable disease) in contrast to patients without amplification. Lack of significant difference in MBC (post-relapse) survival according to c-myc status could be due to a better response of patients to appropriate treatment (chemotherapy). It is possible that negative prognostic impact of c-myc amplification is masked with increased responsiveness to chemotherapy.     

Deficiency of T helper cells in transient hypogammaglobulinemia of infancy

We studied 17 patients with transient hypogammaglobulinemia of infancy to define the immunologic defect responsible for this disorder. The number of circulating B cells in these patients was normal, as was the ability of the B cells to synthesize immunoglobulins when stimulated with Epstein-Barr virus, a direct B-cell activator. However, the capacity of the B cells to synthesize IgG in response to pokeweed mitogen, a T-cell-dependent B-cell activator, was depressed. Experiments with cultured lymphocytes indicated that excess suppressor-cell activity was not present in these patients, but that their T cells were deficient in providing help to B cells from their normal parents. A numerical deficiency in T4-positive (T4+) helper cells was found. Patients who had recovered from the disorder had a normal number of T4+ helper cells. Our results indicate that a numerical, as well as a functional, deficiency in helper T cells underlies the deficiency in IgG production in transient hypogammaglobulinemia of infancy.     

Transient hypogammaglobulinemia of infancy presenting as Staphylococcus aureus sepsis with deep neck infection.

Transient hypogammaglobulinemia of infancy (THI) is characterized by a prolongation and accentuation of the physiologic hypogammaglobulinemia normally occurring during the first 3 to 6 months of life and recovers spontaneously between 18 and 36 months of age. Infants with THI may remain asymptomatic or develop recurrent sinopulmonary infections, but severe or life-threatening infections are rare. We report a case of THI in a previously healthy 1-year-old girl with Staphylococcus aureus sepsis who subsequently developed deep neck infection confirmed by magnetic resonance imaging. Intravenous oxacillin was administered for 21 days and she recovered completely. Immunologic studies were normal except for decreased immunoglobulin G levels. Under the impression of hypogammaglobulinemia with severe infection she received regular intravenous immunoglobulins (IVIG) replacement therapy every 4 weeks. One year later, the immunoglobulin concentrations had returned to the normal range even though IVIG had been discontinued for 4 months. This case report highlights the possibility of severe infection in THI, a disease which usually has a benign clinical course. As the diagnosis of THI can only be made with certainty in retrospect, long-term follow-up of clinical and immune system status is necessary.     

Absent specific viral antibodies in patients with transient hypogammaglobulinemia of infancy

Of 247 patients referred to the Pediatric Immunology Clinic, University of Connecticut Health Center, Farmington, Conn., for recurrent infections, 13 patients were found to have an abnormal delay in the onset of IgG synthesis and prolongation of the physiologic hypogammaglobulinemia of infancy. At first observation, their mean age was 10.6 months. All patients had abnormally low serum IgG levels (less than or equal to 2 SD for their age). The mean serum IgG level for our patient population was 270 +/- 81 mg/dl; the mean serum IgG level of the control group was 749 +/- 440 mg/dl. Clinically, these patients were first observed with recurrent otitis media, respiratory infections, bronchitis and/or asthma, and formula intolerance. Despite recurrent respiratory tract infections, specific antibodies to the respiratory viruses were absent in nine of 11 patients tested who were observed before 17 months of age. On follow-up, two of the 13 patients never developed specific antibodies to viral agents, although their serum IgG levels normalized; one patient became serology positive at the same time that the serum IgG normalized. In two patients, the serum IgG levels returned to within the normal range for age before the appearance of specific viral antibodies. In eight patients, the appearance of specific viral antibodies was detected before the serum IgG levels returned to normal. Five of these eight patients were treated for short periods (9 months) with replacement immune serum globulin (intramuscular) therapy and did clinically well. These observations suggest that replacement immune serum globulin for short periods of time does not appear to suppress the production of specific, naturally occurring antibodies and the resolution of the hypogammaglobulinemia.     

Application of CD27 as a marker for distinguishing human NK cell subsets

It has long been recognized that human NK cells can be dividedinto two phenotypically and functionally distinct subsets, basedon their levels of expression of CD56. We recently found thatCD27 distinguishes subsets of mature mouse NK cells. Here wereport that CD27 can be used as a marker to discriminate humanNK cell subsets. The majority of peripheral blood human NK cellswere CD27^lo/CD56^dim NK cells, whereas the minor CD27^hi NK cellpopulation correspondingly displayed a CD56^bright phenotype.Distinctions between CD27^lo and CD27^hi NK cells in their receptorexpression and typical NK cell functions such as cytotoxicityand cytokine production can be easily delineated. Therefore,we propose the dual use of CD27 and CD56 as maturation/subsetmarkers for human NK cells. The identification of CD27 subsetsin both mice and humans will allow more accurate projectionsof the role of NK cell subsets in murine models of human pathologieswhere NK cells are involved.     

TRKB acts as a prognostic predictive marker in Her-2 positive breast cancer

Human epidermal growth factor receptor 2 (Her-2) positive breast cancer (BC) was associated with an increased risk for brain metastases. Tropomyosin receptor kinase (TRKB) is a specific binding receptor for brain-derived neurotrophic factor associated with brain metastases. However, TRKB was still unknown to be involved in Her-2 positive BC. A tissue microarray comprised of 60 Her-2 positive BC cases and 32 matched adjacent normal samples was analyzed for TRKB expression by immunohistochemical staining. Results were compared to clinicopathologic and survival data by univariate and multivariate analysis. Furthermore, we explored the co-expression genes and related functional proteins using GEPIA, Kaplan-Meier plotter, LinkedOmics and PPI. We found that TRKB protein expression levels were elevated in Her-2 positive BC, and high levels of TRKB expression were associated with vascular invasion, more lymph nodes metastases and more advanced TNM stage as well as poorer OS. TRKB was confirmed as an independent prognostic factor for Her-2 positive BC by univariate and multivariate analysis. Besides, enrichment analyses revealed that protein kinase B signaling was highly correlated to TRKB in Her-2 positive BC. Therefore, TRKB may act as a potential prognostic target and biomarker for Her-2 positive BC.     

The PIWI protein acts as a predictive marker for human gastric cancer

Purpose

To investigate the expression of the human PIWI subfamily proteins in gastric cancer and their potential roles in the occurrence, development and prognosis of gastric cancer.

Methods and patients

Expression of the PIWI proteins were assessed by immunohistochemistry (IHC) in tissue microarrays (TMA), containing paired tumor tissue and adjacent non-cancer tissue from 182 patients who had undergone surgery in hospital for histologically proven gastric cancer (GC). Prognostic value and correlation with other clinicopathologic factors were evaluated in two classifications.

Results

The expression of PIWIL1-4 was significantly higher in tumor tissue than that in adjacent tissue; A significant correlation was observed between the higher expression of PIWI protein with the T stage, lymph node metastasis and clinical TNM (cTNM); Survival analysis by Kaplan-Meier survival curve and log-rank test demonstrated that elevated PIWIL1 and PIWIL2 expression in cancer tissue predicted poorer overall survival (OS) compared with group in lower expression (36.5% VS 67.6%; 37.4% VS 54.2%; respectively). Notably, multivariate analyses by Cox’s proportional hazard model revealed that expression of PIWIL1 was an independent prognostic factor in gastric cancer.

Conclusions

The PIWI subfamily protein is an absolutely key molecular factor along with the tumor occurrence and development. And the PIWI protein could act as a potential biomarker for prognosis evaluation of gastric cancer.     

Topoisomerase II-alfa gene as a predictive marker of response to anthracyclines in breast cancer

Amplification or deletion of the topoisomerase IIα (TOP2A) gene in breast cancer has been related with responsiveness to anthracyclines-based chemotherapy. The purpose of this study was to evaluate the predictive value of TOP2A gene for the efficacy of neo-adjuvant anthracycline in a population with locally advanced breast cancer. Sixty-two patients were included, and the status of TOP2A gene was determined by in situ hybridization method. Treatment efficacy was determined by clinical and pathological response and overall survival. TOP2A gene alterations were found in 22.6% (21.0% of cases with amplification and 1.6% with deletion), and these tumors were biologically more aggressive, with higher nuclear grade, more frequently with HER2 amplification and inflammatory type. Also in these tumors response to chemotherapy appeared to be increased. There was a higher clinical and pathological response rate (complete pathological response of 21.4% vs. 8.3%), a trend toward longer progression-free survival (82.51 vs. 63.12 months) and a trend to increased overall survival (92.08 months; 95% CI 82.81–101.35 vs. 73.40 months; 95% CI 63.44–83.36; p = 0.113). These results corroborate that the TOP2A gene alterations may play an important role in determining anthracycline sensitivity in breast cancer.     

A transgenic marker expressed on discrete populations during B-cell development

We describe a transgenic mouse strain that selectively express a surface marker (huCD25) on transitional B cells, pre-B cells and a lineage unidentified bone marrow (BM) population. We show that a subpopulation of B cells in Peyer's patches, spleen, blood and BM expressed the transgenic huCD25 marker on the cell surface. In the spleen, the huCD25 expression was found on transitional B cells, that had not yet been recruited into the recirculating pool. In the BM a fraction of the B220low surface immunoglobulin (Ig) negative PB493+ pre-B cells were huCD25+. HuCD25 expression was also seen on practically all immature B cells while the mature recirculating B cells did not express huCD25. A huCD25+B220- cell population was also found in the BM that had not rearranged the Ig heavy chain locus and did not express the lineage markers CD3, T-cell receptor (TCR), CD19 and Mac-1. A low expression of CD4 on these cells may indicate that they represent a noncommitted, hematopoetic progenitor cell population.     

The negative as potential prognostic marker of outcome in psychotherapy

From 73 patients, 10 with the best, and 10 with the worst outcomes after psychotherapy were compared statistically for the frequency of ‘not’ and ‘never’ at commencement of psychotherapy, and for change in their frequency after therapy. A highly statistically significantly difference was found at commencement of psychotherapy in that the negative was used more frequently by the worst outcome group (p=.006). No significant differences were found for change in frequency after therapy. Thus, the frequent use of the negative at commencement of psychotherapy seems to predict a poor outcome.     

CINSARC signature as a prognostic marker for clinical outcome in sarcomas and beyond

Prognostication is a key issue for sarcoma patients' care as it triggers the therapeutic approach including chemotherapy, which is still not standard for localized patients. Current prognostic evaluation, based on the FNCLCC grading system, has recently been improved by the CINSARC signature outperforming histology‐based grading system by identifying high‐risk patients in every grade, even in those considered as low. CINSARC is an expression‐based signature related to mitosis and chromosome integrity with prognostic value in a wide range of cancers additional to sarcoma. First developed with frozen material, CINSARC is now coupled with NanoString technology allowing evaluation from FFPE blocks used in clinical practice. Consequently, CINSARC is currently evaluated in clinical trials with a dual objective of demonstrating the benefit of chemotherapy in sarcoma patients and testing its response prediction. Considering its overarching value in oncology, its development is welcome in any cancers where the prognostication needs to be improved.