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Free radicals and scavenging enzymes in chronic tonsillitis.

OBJECTIVE: This study aimed to define the relationship between chronic tonsillitis and levels of malondialdehyde and superoxide dismutase in free radical and antioxidant forms. It is suggested that free oxygen radicals may play a role in chronic tonsillitis. Materials and methods One hundred twenty-four patients were enrolled in the study. Tonsillectomy was performed via the usual dissection-snare method. Venous blood was taken preoperatively and at 2 weeks postoperatively. Blood samples and tonsil specimens were evaluated for malondialdehyde and superoxide dismutase analysis. RESULTS: The levels of malondialdehyde and superoxide dismutase in plasma were compared preoperatively and postoperatively, and there were statistically significant differences between these levels (P < 0.05). In contrast, the levels of malondialdehyde and superoxide dismutase in tonsil tissue were not correlated with the plasma levels of malondialdehyde and superoxide dismutase in pretonsillectomy and posttonsillectomy terms (P > 0.05). CONCLUSION: The presence of malondialdehyde and superoxide dismutase in plasma and tonsil tissue reinforces the involvement of oxidative stress in the pathophysiology of chronic tonsillitis.

Adolescent↗

Distribution and phenotype of Epstein-Barr virus-infected cells in human pharyngeal tonsils.

Although Epstein-Barr virus (EBV) is often found in human tonsils, it remains to be precisely determined in what cells and microenvironment the virus is present. Although generally regarded as a B lymphotropic virus, EBV is associated with non-B-cell tumors, for example, NK/T-cell lymphoma, carcinoma, and leiomyosarcoma. To provide a basis for understanding the origin and biology of EBV-infected non-B cells, the immunophenotype of all EBV-infected cells in reactive human tonsils was determined by subjecting tonsil sections to dual/triple EBER in situ hybridization and immunohistochemistry with monoclonal antibodies to T cells (CD3, CD4, CD8, CCR3), B cells (CD20), plasma cells (CD138), natural killer (NK) cells (PEN5), and epithelial cells (cytokeratin), as well as frozen section immunostaining with antibodies to EBV latent proteins EBNA1, EBNA2, LMP1, and EBV early protein BZLF1. Most tonsils contained nearly equal numbers of EBNA1- and LMP1-positive cells (latency program) while only a few contained EBNA2-positive cells (growth program). More than 1000 EBER-positive cells from six tonsils were detected in the interfollicular zone (59%), tonsillar crypts (26%), and follicles (15%). Most (82%) EBER-positive cells are CD20-positive B cells, 7% are CD3-positive T cells, and 11% are cells of indeterminate lineage, often with plasmacytoid morphology. However, no EBER-positive plasma cells were identified. Rare EBER-positive NK cells and EBER/BZLF1-positive epithelial cells were identified. The direct demonstration of EBV within rare T cells, NK cells, and epithelial cells in reactive human tonsils provide a basis for further understanding of the origin of EBV-associated tumors of non-B-cell type.

Adolescent↗

Association of lip posture and the dimensions of the tonsils and sagittal airway with facial morphology.

The specific contribution of enlarged tonsils or adenoids to craniofacial growth remains unknown, and there is no agreement in the literature as to the significance of lip posture. This study assessed the separate associations of lip posture, sagittal airway size, and tonsil size with selected cephalometric measures. Clinical and cephalometric data of 207 children who presented for evaluation of tonsil and/or adenoid problems were evaluated. Multiple linear regression was used to assess the linear relationship between each of the three parameters and the cephalometric dependent variables. Open lip posture, reduced sagittal airway, and large tonsils were each associated statistically with a characteristic but different skeletal configuration. This association was proportional. Specifically, a more open lip posture was associated with a more backwardly rotated face and larger lower facial height. Reduced sagittal airway size was associated with en bloc backward relocation of the maxilla and mandible. Because the sella-nasion dimension shortened proportionally, the SNA and SNB angles were not affected. Larger tonsils were associated with more forward relocation and rotation of the maxilla and mandible and increased SNA and SNB angles. Because each of the three parameters was associated proportionally with a different craniofacial morphology, it is concluded that lip posture, sagittal airway size, and tonsil size represent three different and unrelated phenomena with respect to their effects on craniofacial growth and form.

Adolescent↗

How well do parents recognize the difference between tonsillitis and other sore throats?

Little evidence exists from randomized controlled trials to support the role of tonsillectomy in children with recurrent tonsillitis. Despite this, parents report a great change for the better in their children following the operation. Previous trials vary in their inclusion criteria, both in terms of the definition of tonsillitis and in the frequency of sore throats required before entry into the study is permitted. The aims of this study are to define tonsillitis from the perspective of parents whose children are awaiting tonsillectomy for recurrent sore throats, and to determine whether such parents have a better understanding of the difference between tonsillitis and other sore throats than parents of children from a normal population control group. These aims demonstrate whether parents who request tonsillectomy for their children do so on the basis of a recognized clinical problem. We report that parents can clearly identify a sore throat illness that they call tonsillitis and which is differentiated from other sore throats by different symptom complexes. The parental history is important in the assessment of a child prior to tonsillectomy. The views of parents whose children have recurrent tonsillitis must be further investigated if the difference between randomised controlled trial results and parents' opinions regarding the benefit of tonsillectomy is to be understood.

Case-Control Studies↗

Analysis of human tonsil and cancer DNAs and RNAs for DNA sequences of group C (serotypes 1, 2, 5, and 6) human adenoviruses.

Group C human adenoviruses (Ads) of serotypes 1, 2, 5, and 6 infect most children and commonly cause latent infections of lymphoid tissues. Ads transform cells into a malignant-like phenotype; the oncogenic genetic information is in the left 8% of the viral genome, in the HindIII-G DNA fragment. We have investigated the molecular basis for group C Ad latent infections in human tonsils as well as whether these viruses are linked to human cancer. Tonsil or cancer DNAs and RNAs were assayed for Ad sequences by liquid-phase saturation-hybridization with in vitro-labeled Ad5 HindIII-G fragment. About 25% of the 52 tonsils analyzed contained DNA or RNA sequences specific to HindIII-G, indicating that Ad transforming sequences are expressed as RNA in tonsils. Southern blotting analysis of four tonsil DNAs revealed multiple copies of the complete Ad genome in a free state and provided evidence for an unusual form of the Ad genome, possibly Ad DNA integrated into cellular DNA. In assays of human cancers, no Ad sequences were detected in DNAs from 26 squamous cell carcinomas (Cas), 3 adenocarcinomas, 4 oat cell Cas, 5 stomach Cas, 5 small intestine Cas, 15 colon Cas, 6 rectum Cas, 5 Hodgkin and 6 non-Hodgkin lymphomas, and 2 breast Cas. Reconstruction experiments indicated that the HindIII-G probe could detect 1 copy per cell of 0.2-0.3% of the viral genome. No HindIII-G-specific sequences were detected in RNAs from 21 squamous cell Cas, 3 oat cell Cas, 2 stomach Cas, or 18 colon Cas. In six other experiments using the complete Ad2 genome as probe, no Ad sequences were found in DNAs from 6 lung Cas, 12 normal lung tissues, 33 gastrointestinal Cas, 19 normal gastrointestinal tissues, 6 Hodgkin lymphomas, 3 breast Cas, or 4 kidney Cas, at a sensitivity of about 1 copy per tumor cell of 5-10% of the Ad2 genome. All Ad-induced cancer cells should contain at least 1 copy of 1-6% of the viral genome, the minimal size of the transforming region, and probably should contain multiple copies of more of the genome. Therefore, our data are definitive evidence against group C Ads being the cause of the cancers tested, which represent about 50% of the cancer incidence in the United States. Of additional interest, we did not detect Ad2 sequences in RNAs from 7 human placentas, 12 normal lungs, or 19 normal gastrointestinal tissues (nor in 44 cancer or 23 tonsil RNAs). Thus, we did not confirm a recent report of the presence of Ad2 RNA in RNAs from human placentas; the possibility that a small population of cells in placenta expresses group C "related" sequences is not ruled out.

Adenoviruses, Human↗

Neutrophils are hyperactive in recurrent tonsillitis.

The pathogenesis of recurrent tonsillitis has not been fully explored. Most studies in this field have focused on pathogenic bacteria whereas less research has been done concerning the host defense. In earlier studies it was shown that there is an active cellular defense in the tonsillar surface secretion, consisting of phagocytes, and therefore in this study the possibility that this defense is altered in patients with recurrent tonsillitis was explored. Neutrophils were obtained from tonsillar surface secretions in eight patients with recurrent tonsillitis and eight healthy volunteers and the capacity of the neutrophils to respond to chemotactic stimuli was examined in an in vitro system. A significantly higher fraction of the neutrophils obtained from the patients with recurrent tonsillitis responded to chemotactic stimuli compared to those obtained from the healthy volunteers. It is concluded that there is a hyperactive cellular defense in the tonsillar surface secretion in patients with recurrent tonsillitis. This finding and its significance in the pathogenesis of recurrent tonsillitis are discussed.

Adolescent↗

Penicillin concentrations in sera and tonsils after intramuscular administration of benzathine penicillin G to children.

BACKGROUND: The optimal regimen of benzathine penicillin G for secondary prevention of rheumatic fever is controversial. Data from serum pharmacokinetic studies do not fully agree on the period of protection after drug administration. Data from concentration of penicillin in tonsils may provide additional information. METHODS: To evaluate penicillin concentrations in palatine tonsils and in sera 1, 10, 14 and 21 days after intramuscular injection of benzathine penicillin G 40,000 IU/kg, 58 children between 4 and 12 years of age with chronic tonsillitis and indication for tonsillectomy were given the study drug 1, 10, 14 or 21 days before surgery. Blood and tonsil samples were obtained during surgery, and penicillin concentrations were determined microbiologically by the agar well diffusion technique. RESULTS: Mean serum penicillin concentrations 1, 10, 14 and 21 days after drug administration were, respectively, 0.080, 0.031, 0.023 and 0.014 microg/ml. Mean penicillin concentrations in tonsils at 1, 10, 14 and 21 days were 0.023, 0.010, 0.007 and 0.002 microg/g, respectively. Detectable penicillin concentration in tonsils (method sensitivity, 0.006 microg/g) was obtained in all patients on Day 1 and in 91% and 83.3% of patients on Days 10 and 14, respectively. By Day 21 this proportion was reduced to 30%. CONCLUSIONS: The results of this study suggest that penicillin values may be inadequate for prevention of rheumatic fever by Week 3 of administration in a majority of children.

Analysis of Variance↗

Biologically active neutrophil chemokine pattern in tonsillitis.

To gain an insight into the mechanisms of chronic and acute inflammation, the production of neutrophil chemokines in different types of tonsillitis - hyperplastic tonsillitis (HT), recurrent tonsillitis (RT) and peritonsillar abscesses (PA) - was investigated. The chemokines interleukin-8 (IL-8), growth-related oncogene-alpha (GRO-alpha), epithelial cell-derived neutrophil attractant-78 (ENA-78) and granulocyte chemotactic protein-2 (GCP-2) were detected and shown to have different biological activities. With respect to the biological properties of CXC chemokines, the biological activity of the chemokines was identified using a three-step high-performance liquid chromatography (HPLC) technique, a bioassay involving measurement of neutrophil chemotaxis in a single Boyden chamber in tissue of HT, RT and PA. Using reverse transcription-polymerase chain reaction (RT-PCR), the chemokine concentrations were determined in the different tonsillitis entities. The chemokine pattern was dominated in PA by IL-8 and GRO-alpha and in RT by GRO-alpha. Hyperplastic tonsils of patients without a history of infection generated about five times lower IL-8 than PA. A protein concentration of GCP-2 was induced in PA and RT, whereas ENA-78 remained the same in all entities. In conclusion, it would appear that IL-8 was up-regulated in acute inflammation, whereas GRO-alpha dominated in chronic inflammation. ENA-78 seems not to play a pivotal role in inflammatory processes in tonsils. GCP-2 may serve as a substitute chemokine in certain inflammatory conditions as its quantity of mRNA and protein was higher in RT and PA than in HT.

Acute Disease↗

Neutrophil chemokines in epithelial inflammatory processes of human tonsils.

CXC chemokines are thought to play an important role at sites of inflammation. Because ELR(+) CXC chemokines are expressed in different types of tonsillitis we investigated the role of the surface/crypt epithelium of human tonsils in producing ELR(+) CXC chemokines: interleukin (IL)-8 (CXCL8), ENA-78 (CXCL5), GRO-alpha (CXCL1) and GCP-2 (CXCL6). Tonsillar tissue was obtained from patients undergoing tonsillectomy and chemokine expression was investigated by means of immunohistochemistry. A549 cells were used as a model to study kinetics of chemokine expression in epithelial cells. Cells were stimulated with tumour necrosis factor (TNF)-alpha, lipopolysaccharide (LPS) and supernatants derived from aerobic/anaerobic Staphylococcus aureus strains. Chemokine expression was measured by quantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). We observed epithelial expression of IL-8, GRO-alpha and GCP-2 in different types of tonsillitis, whereas ENA-78 was rarely expressed. In A549 cells abundant expression of ENA-78 was detected. IL-8 and GCP-2 are expressed in an acute type of tonsillitis whereas GRO-alpha was frequently detectable both in chronically and acutely inflamed tonsils. ENA-78 does not seem to play a pivotal role in tonsillitis in vivo.

Adolescent↗

Economic benefit of tonsillectomy in adults with chronic tonsillitis.

To determine the economic impact of adult chronic tonsillitis and the economic improvement from adult tonsillectomy, we studied patients who underwent adult tonsillectomy for chronic tonsillitis with the Glasgow Benefit Inventory and a questionnaire for disease severity parameters before and after tonsillectomy. The economic impact of chronic tonsillitis and adult tonsillectomy were computed with a break-even time analysis model. Eighty-three adult patients (average age, 27.3 years) completed the study with an average duration of follow-up of 37.7 months. The patients exhibited a mean improvement in quality of life of +27.54 +/- 4.63 after tonsillectomy according to the Glasgow Benefit Inventory. In the 12 months following the procedure, tonsillectomy resulted in yearly mean decreases in number of weeks on antibiotics by 5.9 weeks, number of workdays missed because of tonsillitis by 8.7 days, and physician visits for tonsillitis by 5.3 visits. In considering the medical costs of tonsillectomy only, the break-even point was found to be 12.7 years, whereas considering the overall economic impact of tonsillectomy resulted in a break-even point of 2.3 years after the procedure. We conclude that tonsillectomy results in significant improvement in quality of life, decreases health-care utilization, and diminishes the economic burden of chronic tonsillitis in the adult patient population.

Adult↗

On the treatment of recurrent tonsillitis by vaccination.

This paper reports on the experience of using vaccination in the treatment of recurrent tonsillitis. All of the 82 patients, who were aged between 5 and 67 years and who suffered from recurrent tonsillitis, were given an injection of a bacterial vaccine (Broncasma Berna, Swiss Serum Institute, Berne) that has been used in many countries for three decades. A dose of 0.05 ml was given once every 4 to 14 days (average once a week) on 3 to 20 occasions (average 8). The treatment prevented 75 of the 82 patients (91%) from having attacks of recurrent tonsillitis for extended periods. These 75 patients included three who relapsed into recurrent tonsillitis in spite of having tonsillectomies. As the dose used was relatively small, none of the patients complained of spontaneous pain or fever although three complained of slight tenderness at the injection point on the following day. The effect on recurrent tonsillitis of the Paspat intracutaneous injection (Luitpold Werk, Munich) has also been investigated recently. It was effective in two patients who did not respond to Broncasma Berna. In this retrospective study, low doses of bacterial vaccine reduced the incidence of recurrent tonsillitis and were well tolerated.

Adolescent↗

In vivo migration of tonsil lymphocytes in rheumatoid synovial tissue engrafted in SCID mice: involvement of LFA-1.

Integrin-adressin binding is a critical step in lymphocte attachment to target tissues. The mucosal recognition systems (alpha E beta 7, alpha 4 beta 7, MADcam-1) have been implicated in the autoimmune process in rheumatoid arthritis. We developed a model for in vivo study of radio-labelled lymphocyte circulation and their attachment to human rheumatoid synovium. We studied the homing of tonsil lymphocytes, considered as mucosal lymphocytes, and the involvement of alpha E beta 7 integrin and LFA1 in the homing of tonsil lymphocytes. We engrafted human rheumatoid synovium subcutaneously in 6 week old SCID CB17 mice. Three weeks later, we injected intraperitoneally 20 IO6 human peripheral blood or tonsil mononuclear cells, previously labelled with 3 mCFi HMPAO-99mTc. A mouse total body scintigram was obtained 20 h postinjection. The same protocol was performed after treatment of the MNC and mAb against LFA-1 (CD11a) or alpha E beta 7 (CD103). Tonsil MNC retention in the rheumatoid synovial graft 20 h post-injection was enhanced compared to blood MNC (12731 +/- 8297cpm/200 pixel) versus 5982 +/- 4713cpm/200 pixel, p < 0.05). A monoclonal antibody against LFA 1 decreased the activity in the graft (4152 +/- 1287 cpm/200 pixel), p < 0.05. No significant difference in tonsil MNC attachment to rheumatoid synovial tissue was observed with a mAb against alpha E beta 7 (8057 +/- 5009 cpm/200 pixel). Our results showed an increase in radiolabelled mucosal MNC migration in synovial tissue engrafted in SCID mice compared with blood MNC. Moreover, the date suggest that LFA-1 but not the alpha E beta 7 integrin is involved in tonsil MNC binding to synovial tissue in RA.

Animals↗

[Induction and functional analysis of gamma delta TCR-bearing T cells from human tonsil by Streptococcus pyogenes stimulation].

The killer cell characteristics of gamma delta TCR-bearing T cells induced from human tonsil by streptococcus pyogenes stimulation were examined. Immunohistologic staining of tonsil showed that gamma delta TCR-bearing T cells were mainly located in the interfollicular area connected with stratified squamous epithelium. Streptococcus pyogenes could induce the proliferation of gamma delta TCR-bearing T cells from tonsil in the presence of low-dose of IL-2. This streptococcus pyogenes-induced gamma delta TCR-bearing T cell proliferation was likely to be independent on the IL-2/IL-2R system since an obvious inhibition was not observed with anti-IL-2 and anti-IL-2R mAbs. More importantly, these gamma delta TCR-bearing T cells exhibited cell-mediated cytotoxic activity in a 4 hr 51 Cr-release assay. In addition, immunocytochemical staining revealed that these cells contained a killer protein perforin. These results demonstrate that gamma delta TCR-bearing T cells from tonsil exhibit typical killer cell characteristics. These data also suggest that gamma delta TCR-bearing T cells in human tonsil may play a cytotoxic role in protecting the integrity of tonsil from infection.

Cytotoxicity, Immunologic↗

RAG1 and RAG2 expression by B cell subsets from human tonsil and peripheral blood.

It has been suggested that B cells acquire the capacity for secondary V(D)J recombination during germinal center (GC) reactions. The nature of these B cells remains controversial. Subsets of tonsil and blood B cells and also individual B cells were examined for the expression of recombination-activating gene (RAG) mRNA. Semiquantitative analysis indicated that RAG1 mRNA was present in all tonsil B cell subsets, with the largest amount found in naive B cells. RAG2 mRNA was only found in tonsil naive B cells, centrocytes, and to a lesser extent in centroblasts. Neither RAG1 nor RAG2 mRNA was routinely found in normal peripheral blood B cells. In individual tonsil B cells, RAG1 and RAG2 mRNAs were found in 18% of naive B cells, 22% of GC founder cells, 0% of centroblasts, 13% of centrocytes, and 9% of memory B cells. Individual naive tonsil B cells containing both RAG1 and RAG2 mRNA were activated (CD69(+)). In normal peripheral blood approximately 5% of B cells expressed both RAG1 and RAG2. These cells were uniformly postswitch memory B cells as documented by the coexpression of IgG mRNA. These results indicate that coordinate RAG expression is not found in normal peripheral naive B cells but is up-regulated in naive B cells which are activated in the tonsil. With the exception of centroblasts, RAG1 and RAG2 expression can be found in all components of the GC, including postswitch memory B cells, some of which may circulate in the blood of normal subjects.

Adult↗

Yersinia pseudotuberculosis with limited genetic diversity is a common finding in tonsils of fattening pigs.

A total of 425 pig tonsils, including 210 tonsils from fattening pigs and 215 from sows, from seven different abattoirs in Finland were studied for the occurrence of Yersinia pseudotuberculosis from 1999 to 2000. The mean prevalence of Y. pseudotuberculosis in fattening pig tonsils was 4%, varying from 0 to 10% between slaughterhouses. Y. pseudotuberculosis was not recovered from sow tonsils. All 30 Y. pseudotuberculosis isolates from eight pig tonsils were recovered after cold enrichment. Seventeen isolates from seven tonsils were found after cold enrichment for 14 days, followed by alkali treatment. Y. pseudotuberculosis was not isolated after direct plating, overnight enrichment, or selective enrichment. All 30 isolates belonged to bioserotype 2/0:3 and carried the virF gene in the virulence plasmid. The isolates exhibited calcium dependence and Congo red absorption. The pyrazinamidase test gave variable results. All isolates were characterized with pulsed-field gel electrophoresis (PFGE). Using SpeI, NotI, and XbaI enzymes, seven, five, and two different PFGE patterns were obtained, respectively. A total of 11 genotypes, gI to gXI, identified by a combination of the various SpeI, NotI, and XbaI profiles, were detected. Three pigs were found to carry more than one genotype. Overall, variations between PFGE patterns were small, indicating genetic homogeneity among pig strains of bioserotype 2/0:3.

Abattoirs↗

Presence of Yersinia enterocolitica in tissues of orally-inoculated pigs and the tonsils and feces of pigs at slaughter.

In order to study the early events associated with infection of swine by Yersinia enterocolitica, 42 five-week-old crossbred piglets were inoculated per os with approximately 10(8) Y. enterocolitica O:3. Groups of 5 animals (and one negative control) were euthanized 30 min, 3, 6, 12, 24, 48 and 72 h following the infection. Palatine tonsils, retropharyngeal and mesenteric lymph nodes, esophagus, duodenum, jejunum, ileum (and Peyer's patches), stomach, liver, spleen and feces (from colon) were collected and analyzed for the presence of Y. enterocolitica by standard bacteriological procedures. Natural infections were also analyzed, as a complementary study, by taking one-gram samples of fecal material and tonsils from 291 pig carcasses less than 3 h after slaughter and culturing them for Y. enterocolitica using a cold enrichment technique. Within 30 min, Yersinia enterocolitica O:3 was already present at most sites. The presence of Y. enterocolitica in the liver of 3 out of 10 animals and also in the spleen of 3 out of 10 piglets, within the first 3 h postinfection, but not at later times (with one exception), probably indicated a transient bacteremia accompanying the initial stages of infection. The tonsils were colonized in most animals (13/20) as the bacteria remained present from 12 to 72 h postinfection, while only 4 out of 20 fecal samples were found to be positive over the same period. Up to 10(4) colony-forming units of Y. enterocolitica per gram of tonsil and fecal material were recovered. Finally, among the 291 animals sampled at the abattoir, a total of 79 were found positive, 70 of the tonsils sampled were positive, and bacteria were recovered in 17 fecal samples. It is therefore suggested that palatine tonsils are the most reliable tissue for the indication of an infection/colonization by Y. enterocolitica O:3 in swine and that the removal of this tissue during the slaughter process should be considered in order to minimize the possibility of contamination of meat products.

Abattoirs↗

[Evaluation of the function of palatine tonsil with regard to phagocytosis].

Trypan blue and Indian ink made in Fuzhou as vital staining dyes were injected into the abdominal cavities of rabbits. Nine days later, the tonsil, thymus, spleen and lymph nodes were examined. Numerous pigment granules were observed in the macrophages of the spleen and lymph node. However, no phagocytosis of pigment granules was found in the palatine tonsil and thymus. It indicated that the function of palatine tonsil as a lymph organ is to contact exogenous antigens and to take part in immune response. It is important for children, especially infants, to establish a complete immune mechanism. The Tonsils reach their maximum development in childhood and thereafter decline. Once the palatine tonsils become chronic foci, surgical removal of the diseased tonsils is necessary.

Animals↗

Association between tonsil size and immunoglobulin concentrations.

Tonsil size and serum immunoglobulin (G, A, M and D) measurements were studied in 1049 individuals during a health survey on the West Coast of Newfoundland. The data were subjected to multifactorial analysis of variance. People with vestigial tonsils had the lowest mean IgG-levels, whereas those with naturally absent tonsils had the highest mean IgG-levels. There were also statistical differences in the mean IgA levels in relation to tonsil size. Individuals with vestigial tonsil sizes had the highest mean IgA levels whereas those who had had a tonsillectomy had the lowest. There are no tonsil size-related differences for IgM or IgD concentrations.

Female↗