ABSTRACT
Cryptococcal infection of central nervous system commonly involves meningitis or meningoencephalitis, but rarely mimics inflammatory myelitis. We present short segment myelitis as a dominant manifestation caused by Cryptococcus neoformans in a patient with nephrotic syndrome under immunosuppressive therapy. This case report highlights Cryptococcus neoformans as a potential etiological factor for short segment myelitis in immunocompromised hosts.
Subject(s)
Cryptococcosis , Cryptococcus neoformans , Myelitis , Humans , Myelitis/microbiology , Myelitis/drug therapy , Myelitis/diagnosis , Cryptococcosis/drug therapy , Cryptococcosis/microbiology , Cryptococcosis/diagnosis , Cryptococcus neoformans/isolation & purification , Male , Immunocompromised Host , Middle Aged , Nephrotic Syndrome/complications , Immunosuppressive Agents/therapeutic use , Magnetic Resonance ImagingABSTRACT
BACKGROUND: Cryptococcosis is an infectious disease caused by encapsulated heterobasidiomycete yeasts. As an opportunistic pathogen, cryptococcal inhalation infection is the most common. While Primary cutaneous cryptococcosis is extremely uncommon. CASE PRESENTATION: A 61-year-old woman with a history of rheumatoid arthritis on long-term prednisone developed a red plaque on her left thigh. Despite initial antibiotic treatment, the erythema worsened, leading to rupture and fever. Microbiological analysis of the lesion's secretion revealed Candida albicans, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus epidermidis. Skin biopsy showed thick-walled spores, and culture confirmed primary cutaneous infection with Cryptococcus neoformans. Histopathological stains were positive, and mass spectrometry identified serotype A of the pathogen. The patient was treated with oral fluconazole and topical nystatin, resulting in significant improvement and near-complete healing of the skin lesion within 2.5 months. CONCLUSIONS: Primary cutaneous cryptococcosis was a primary skin infection exclusively located on the skin. It has no typical clinical manifestation of cutaneous infection of Cryptococcus, and culture and histopathology remain the gold standard for diagnosing. The recommended medication for Primary cutaneous cryptococcosis is fluconazole. When patients at risk for opportunistic infections develop skin ulcers that are unresponsive to antibiotic, the possibility of primary cutaneous cryptococcosis needs to be considered.
Subject(s)
Antifungal Agents , Cryptococcosis , Cryptococcus neoformans , Fluconazole , Humans , Female , Middle Aged , Cryptococcus neoformans/isolation & purification , Cryptococcus neoformans/drug effects , Cryptococcosis/drug therapy , Cryptococcosis/microbiology , Cryptococcosis/diagnosis , Cryptococcosis/pathology , Antifungal Agents/therapeutic use , Fluconazole/therapeutic use , Dermatomycoses/drug therapy , Dermatomycoses/microbiology , Dermatomycoses/diagnosis , Dermatomycoses/pathology , Skin/pathology , Skin/microbiology , Treatment Outcome , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/complicationsABSTRACT
OBJECTIVE: Cryptococcosis predominantly presents as a meningoencephalitis in Thailand. Early and expeditious diagnosis is essential for reducing both mortality and morbidity associated with cryptococcal meningitis. We aim to define and establish the diagnostic performances between the benchmark commercially available diagnostic kit (CrAg® LFA) and the large-scale prototype of an inexpensive in-house immunochromatographic test (ICT) based on monoclonal antibody (MAb) 18B7. METHODS: We have developed the large-scale prototype for the rapid detection of cryptococcal polysaccharide antigens by utilizing a single antibody sandwich ICT format employing MAb 18B7, which is highly specific to Cryptococcus neoformans glucuronoxylomannan (GXM) antigens. An in-house MAb18B7 ICT was manufactured in accordance with industry standards under the control of the International Organization for Standardization (ISO) 13485. RESULTS: The diagnostic sensitivity, specificity, and accuracy for the in-house MAb 18B7 ICT were 99.10%, 97.61%, and 97.83%, respectively. The agreement kappa (κ) coefficient was 0.968 based on the retrospective evaluation of 580 specimens from patients living in northern Thailand with clinically suspected cryptococcosis. CONCLUSION: The data suggest that this in-house MAb 18B7 ICT will be highly beneficial for addressing the issue of cryptococcal infection in Thailand. Moreover, it is anticipated that this inexpensive ICT can play a pivotal role in various global strategies aimed at eradicating cryptococcal meningitis among individuals living with HIV by 2030.
Subject(s)
Antibodies, Monoclonal , Antigens, Fungal , Chromatography, Affinity , Cryptococcosis , Cryptococcus neoformans , Sensitivity and Specificity , Humans , Thailand , Antibodies, Monoclonal/immunology , Chromatography, Affinity/methods , Cryptococcosis/diagnosis , Cryptococcus neoformans/immunology , Cryptococcus neoformans/isolation & purification , Antigens, Fungal/analysis , Antigens, Fungal/immunology , Retrospective Studies , Antibodies, Fungal/blood , Polysaccharides/analysis , Polysaccharides/immunology , Male , Female , Adult , Diagnostic Tests, Routine/methods , Middle Aged , Aged , Young AdultABSTRACT
BACKGROUND: Cryptococcosis is a life-threatening disease caused by Cryptococcus neoformans or C. gattii. Neutralizing autoantibodies (auto-Abs) against granulocyte-macrophage colony-stimulating factor (GM-CSF) in otherwise healthy adults with cryptococcal meningitis have been described since 2013. We searched for neutralizing auto-Abs in sera collected from Colombian patients with non-HIV-associated cryptococcosis in a retrospective national cohort from 1997 to 2016. METHODS: We reviewed clinical and laboratory records and assessed the presence of neutralizing auto-Abs against GM-CSF in 30 HIV negative adults with cryptococcosis (13 caused by C. gattii and 17 caused by C. neoformans). RESULTS: We detected neutralizing auto-Abs against GM-CSF in the sera of 10 out of 13 (77%) patients infected with C. gattii and one out of 17 (6%) patients infected with C. neoformans. CONCLUSIONS: We report eleven Colombian patients diagnosed with cryptococcosis who had auto-Abs that neutralize GM-CSF. Among these patients, ten were infected with C. gattii and only one with C. neoformans.
Subject(s)
Antibodies, Neutralizing , Autoantibodies , Cryptococcosis , Cryptococcus gattii , Cryptococcus neoformans , Granulocyte-Macrophage Colony-Stimulating Factor , Humans , Granulocyte-Macrophage Colony-Stimulating Factor/immunology , Autoantibodies/blood , Autoantibodies/immunology , Male , Colombia , Female , Adult , Cryptococcus gattii/immunology , Middle Aged , Cryptococcus neoformans/immunology , Cryptococcosis/immunology , Cryptococcosis/diagnosis , Antibodies, Neutralizing/blood , Antibodies, Neutralizing/immunology , Retrospective Studies , HIV Seronegativity/immunology , Young Adult , AgedABSTRACT
BACKGROUND Cryptococcosis is an invasive fungal infection caused by Cryptococcus species complex. C. neoformans is one of the pathogenic species within the genus. C. neoformans infections often present as an opportunistic infection in severely immunocompromised individuals. Infection of the pericardium in the setting of liver failure is uncommon. We present a case of cryptococcal pericarditis in a patient with liver failure. CASE REPORT A 47-year-old man with a past medical history of psoriatic arthritis, and alcohol use disorder presented to the emergency department with a 2-week history of progressively worsening generalized weakness, malaise, and yellowish skin changes. Physical examination revealed scleral icterus, jaundiced skin, and ascites. Initial laboratory workup revealed thrombocytopenia, transaminitis (aspartate transaminase (AST) level of 502 IU/L, alanine transaminase (ALT) level of 82 IU/L), hyperbilirubinemia (total bilirubin of 15.7 mg/dL), International Nationalized Ratio (INR) of 3.6, and lactic acidosis (lactic acid of 11.7 mmol/L). The patient developed encephalopathy and acute hypoxic respiratory failure requiring intubation. A bedside point-of-care cardiac ultrasound, performed following intubation, revealed a pericardial effusion without signs of tamponade. This finding was later confirmed by a formal transthoracic echocardiogram. Percutaneous pericardiocentesis was performed, and the pericardial fluid culture revealed the presence of C. neoformans. Human immunodeficiency virus (HIV) tests were negative. The patient received antifungal therapy. Due to his poor prognosis, he was transitioned to comfort care and eventually died. CONCLUSIONS This case report describes an unusual presentation of acute liver failure complicated by cryptococcal pericarditis, emphasizing the importance of considering atypical fungal infections in such patients.
Subject(s)
Cryptococcosis , Pericarditis , Humans , Male , Middle Aged , Cryptococcosis/diagnosis , Cryptococcosis/complications , Pericarditis/microbiology , Pericarditis/diagnosis , Liver Failure/etiology , Cryptococcus neoformans/isolation & purification , Antifungal Agents/therapeutic use , Immunocompromised Host , Fatal OutcomeABSTRACT
BACKGROUND Cryptococcosis is an opportunistic fungal infection that typically occurs in patients with compromised immune systems, primarily affecting the respiratory and central nervous systems. However, cryptococcal osteomyelitis is a rare manifestation of cryptococcal infection, characterized by nonspecific clinical features. Here, we present a case of vertebral cryptococcal osteomyelitis in a middle-aged woman and discuss diagnostic approaches. CASE REPORT A 56-year-old woman presented with lower back pain and limited mobility, without fever, and with a history of pulmonary tuberculosis. Physical examination revealed enlarged lymph nodes and tenderness in the thoracic vertebrae. A computed tomography-guided biopsy confirmed granulomatous inflammation caused by Cryptococcus, with abundant 10 µm spherical microbial spores. After 4 weeks of treatment with amphotericin B and fluconazole, symptoms and lesions improved. Upon discharge, the patient was prescribed oral fluconazole. Follow-up examinations showed a stable condition and a negative serum cryptococcal capsular polysaccharide antigen test. CONCLUSIONS Given the rarity and lack of specificity of clinical features of cryptococcal spondylitis, clinicians encountering similar presentations should consider tuberculous spondylitis and spinal tumors as differential diagnoses. Additionally, tissue biopsy of the affected vertebral bodies should be performed early to establish the type of vertebral infection, aiding in diagnosis, treatment, and prognosis.
Subject(s)
Cryptococcosis , Osteomyelitis , Tuberculosis, Spinal , Humans , Female , Middle Aged , Osteomyelitis/diagnosis , Osteomyelitis/microbiology , Osteomyelitis/drug therapy , Cryptococcosis/diagnosis , Cryptococcosis/drug therapy , Diagnosis, Differential , Tuberculosis, Spinal/diagnosis , Thoracic Vertebrae , Antifungal Agents/therapeutic use , Fluconazole/therapeutic use , Tomography, X-Ray ComputedABSTRACT
BACKGROUND: Cryptococcosis is a global invasive mycosis associated with significant morbidity and mortality. Cryptococcal antigen (CrAg) testing from serum and cerebrospinal fluid (CSF) has been regarded as a gold standard for early diagnosis. This study aimed to develop and validate a rapid and sensitive sandwich chemiluminescent magnetic microparticle immunoassay (CMIA) for quantitative detection of CrAg in sera. RESEARCH DESIGN AND METHODS: CMIA is based on magnetic beads modified with capture antibodies and biotinylated antibodies and Streptavidin-polyHRP, where biotinylated antibodies functioned as the recognition element and Streptavidin-polyHRP as the signal component. Assay parameters were first optimized, and then assay performances were evaluated. RESULTS: Under optimized conditions, the total runtime of the CMIA was 22 min. The assay had a wide linear range (2 -10,000 ng/mL) and high analytical sensitivity (0.24 ng/mL), together with acceptable reproducibility, accuracy, and stability. Besides, it exhibited no cross-reactivity with other pathogens. Importantly, the assay showed 92.91% (95% CI, 80.97-93.02%) overall qualitative agreement with a commercial ELISA kit in a retrospective cohort of 55 cases with confirmed cryptococcal infection, and 72 controls without evidence of invasive fungal disease (IFD). CONCLUSION: These results demonstrated that the present study paved a novel strategy for reliable quantitative detection of CrAg in sera.
Subject(s)
Antigens, Fungal , Cryptococcosis , Luminescent Measurements , Humans , Antigens, Fungal/blood , Antigens, Fungal/immunology , Luminescent Measurements/methods , Immunoassay/methods , Cryptococcosis/diagnosis , Cryptococcosis/blood , Reproducibility of Results , Sensitivity and Specificity , Female , Male , Middle Aged , Adult , Cryptococcus/immunology , Retrospective StudiesABSTRACT
A 2-year-old Norwegian Forest cat was presented for evaluation of bilateral purulent nasal discharge and stertorous breathing. A computed tomography (CT) scan of the head revealed an intranasal mass of the left nasal cavity extending behind the tube openings and completely obstructing the nasopharynx. Rhinoscopy confirmed a pinkish, shiny mass. CT scan showed both compartments of the right middle ear filled with abnormal soft tissue attenuating material. There was no change in the bony outline of the middle ear. In the endoscopic examination, after endoscopically assisted tympanocentesis, this material in the accessible dorsolateral compartment proved to be classic polypous tissue in addition to highly viscous glue-like secretions. A secondary otitis media due to a drainage disorder was suspected.Using an endoscopic-interventional approach through the nostril, the nasopharyngeal mass was removed for histopathological examination, in order to restore the nasal airway, and to allow tube drainage. In contrast to cats with classical malignant nasal cavity masses, the cat showed several attachment points of the mass and multiple undulating elevations bilaterally in the nasopharyngeal mucosa.Cytological and histopathological examination identified the mass as a fungal granuloma in the context of a cryptococcus infection only rarely observed in Germany. Molecular genetic analysis confirmed an infection with Cryptococcus neoformans var. grubii.A single intranasal and nasopharyngeal endoscopic debridement resulted in a significant improvement of the clinical signs and a complete healing of the right middle ear (including the tympanic membrane) within 14 days, but not in a complete cure of the disease. The cat was therefore treated with oral itraconazole solution for several weeks.The case report shows that nasal cryptococcosis can also affect cats in Germany. Rhinoscopy reveals a nasopharyngeal mass with multiple attachment points, which is unusual for a neoplasia. In addition to the recommended removal of the mass, oral administration of systemic antimycotics is strongly advised.
Subject(s)
Cat Diseases , Cryptococcosis , Animals , Cats , Cat Diseases/diagnosis , Cat Diseases/microbiology , Cat Diseases/pathology , Cryptococcosis/veterinary , Cryptococcosis/diagnosis , Cryptococcosis/microbiology , Cryptococcosis/pathology , Cryptococcosis/drug therapy , Diagnosis, Differential , Nasopharyngeal Neoplasms/veterinary , Nasopharyngeal Neoplasms/diagnosis , Germany , Tomography, X-Ray Computed/veterinary , Nasopharyngeal Diseases/veterinary , Nasopharyngeal Diseases/diagnosis , Nasopharyngeal Diseases/microbiology , Nasopharyngeal Diseases/pathologyABSTRACT
BACKGROUND: In July 2023, our hospital confirmed one case of lumbar spine infected complicated by Mycobacterium tuberculosis and Cryptococcus neoformans. The patient was admitted due to lower back pain for 1 year and a hard lump for 3 months. Symptoms and signs: Dressing can be seen fixed on the lower back, with severe bleeding. When the dressing is removed, a hard and protruding lump with a size of 6 cm x 8 cm, a sinus tract can be seen near the mass, with a slightly red wound and a sinus depth of about 3 cm. Light red fluid can be seen flowing out. There are no symptoms such as redness, swelling, or heat in the rest of the lower back, and the patient has no other underlying diseases or surgical history. METHODS: Lumbar magnetic resonance imaging and lumbar CT examination; Percutaneous puncture lumbar vertebral biopsy was performed, and the biopsy tissue was subjected to pathological examination, mNGS (metagenomic next-generation sequencing), and acid-fast staining; Extract pus from the lump for fungal culture and ink staining, and identify the fungi through MALDI-TOF MS. RESULTS: Bone destruction and bone marrow edema in the L5 vertebral body, compression of the spinal canal at the L5 vertebral body level; The pathological results of the biopsy tissue indicate granulomatous lesions. The acid-fast staining of the tissue is positive, and the mNGS of the tissue indicates infection with Mycobacterium tuberculosis. A single fungus was cultured from pus and identified by MALDI-TOF MS as Cryptococcus neoformans. Clinically, isoniazid 0.3 g ivgtt + rifampicin 0.45 g qd po + ethambutol 0.25 g qd po + pyrazinamide 0.75 g qd po + fluconazole 0.3 g qd po was administered for treatment. After 11 days, there was slight pain at the incision site, and the original symptoms were significantly relieved. The wound dressing was fixed in place, dry and without obvious exudation. Improved and discharged, followed up for 3 months with no recurrence of the lesion. CONCLUSIONS: mNGS is an effective identification technique that can be used to accurately diagnose suspected infection cases. MALDI-TOF MS has significant advantages over traditional detection methods in shortening detection time. This case achieved satisfactory treatment results for patients through a reasonable treatment plan, which is of great significance for exploring the diagnosis and treatment of similar disease infections.
Subject(s)
Cryptococcosis , Cryptococcus neoformans , Lumbar Vertebrae , Mycobacterium tuberculosis , Humans , Cryptococcus neoformans/isolation & purification , Lumbar Vertebrae/microbiology , Mycobacterium tuberculosis/isolation & purification , Cryptococcosis/diagnosis , Cryptococcosis/microbiology , Cryptococcosis/drug therapy , Male , Tuberculosis, Spinal/diagnosis , Tuberculosis, Spinal/microbiology , Magnetic Resonance Imaging , Antitubercular Agents/therapeutic use , Antitubercular Agents/administration & dosage , Middle Aged , Tomography, X-Ray ComputedABSTRACT
ABSTRACT: This article reports an elderly male patient with nodules and ulcers on the face and behind the left ear after trauma. Primary cutaneous cryptococcosis was confirmed using pathological biopsy, special staining, tissue culture, and fungal sequencing. The patient received a therapeutic intervention involving the administration of the antifungal agent itraconazole. Substantial amelioration of cutaneous manifestations was observed after a 3-month course of treatment. After an elapsed interval, the patient was diagnosed with esophageal tumor. Moreover, the literature on 33 patients with primary cutaneous cryptococcosis published in the past 10 years was also reviewed.
Subject(s)
Antifungal Agents , Cryptococcosis , Dermatomycoses , Humans , Cryptococcosis/drug therapy , Cryptococcosis/pathology , Cryptococcosis/microbiology , Cryptococcosis/diagnosis , Male , Antifungal Agents/therapeutic use , Dermatomycoses/drug therapy , Dermatomycoses/microbiology , Dermatomycoses/pathology , Dermatomycoses/diagnosis , Aged , Itraconazole/therapeutic use , Esophageal Neoplasms/pathology , Esophageal Neoplasms/microbiology , Esophageal Neoplasms/drug therapy , Treatment Outcome , Biopsy , Cryptococcus neoformans/isolation & purificationABSTRACT
Cryptococcus neoformans is a fungal pathogen that causes cryptococcosis primarily in immunocompromised patients, such as those with HIV/AIDS. One survival mechanism of C. neoformans during infection is melanin production, which catalyzed by laccase and protects fungal cells against immune attack. Hence, the comparative assessment of laccase activity is useful for characterizing cryptococcal strains. We serendipitously observed that culturing C. neoformans with food coloring resulted in degradation of some dyes with phenolic structures. Consequently, we investigated the color changes for the food dyes metabolized by C. neoformans laccase and by using this effect explored the development of a colorimetric assay to measure laccase activity. We developed several versions of a food dye-based colorimetric laccase assay that can be used to compare the relative laccase activities between different C. neoformans strains. We found that phenolic color degradation was glucose-dependent, which may reflect changes in the reduction properties of the media. Our food color-based colorimetric assay has several advantages, including lower cost, irreversibility, and not requiring constant monitoring , over the commonly used 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay for determining laccase activity. This method has potential applications to bioremediation of water pollutants in addition to its use in determining laccase virulence factor expression.IMPORTANCECryptococcus neoformans is present in the environment, and while infection is common, disease occurs mostly in immunocompromised individuals. C. neoformans infection in the lungs results in symptoms like pneumonia, and consequently, cryptococcal meningitis occurs if the fungal infection spreads to the brain. The laccase enzyme catalyzes the melanization reaction that serves as a virulence factor for C. neoformans. Developing a simple and less costly assay to determine the laccase activity in C. neoformans strains can be useful for a variety of procedures ranging from studying the relative virulence of cryptococci to environmental pollution studies.
Subject(s)
Cryptococcus neoformans , Food Coloring Agents , Laccase , Humans , Colorimetry/methods , Cryptococcosis/microbiology , Cryptococcosis/diagnosis , Cryptococcus neoformans/enzymology , Food Coloring Agents/metabolism , Fungal Proteins/metabolism , Laccase/metabolism , Phenols/metabolismABSTRACT
We discuss a rare instance of cryptococcoma caused by Cryptococcus gattii in a 55-year-old woman initially treated for suspected COVID bronchopneumonia. The diagnosis posed a challenge due to vague symptoms and unclear imaging findings suggesting malignancy. Postoperative samples confirmed the presence of Cryptococcus gattii through culture of brain tissue and blood. Appropriate therapy was initiated, but despite treatment, it led to a fatal outcome. The case emphasizes the crucial role of microbiologist in early diagnosis of fungal infections of Central Nervous System. Additionally, the delayed diagnosis in immunocompetent individuals highlights the critical need for early recognition and intervention to mitigate potentially fatal outcomes.
Subject(s)
Cryptococcosis , Cryptococcus gattii , Glioblastoma , Humans , Female , Middle Aged , Cryptococcus gattii/isolation & purification , Cryptococcosis/diagnosis , Cryptococcosis/microbiology , Glioblastoma/diagnosis , Diagnosis, Differential , Fatal Outcome , Brain/pathology , Brain/diagnostic imaging , Brain/microbiology , Brain Neoplasms/diagnosis , Antifungal Agents/therapeutic use , COVID-19/diagnosisABSTRACT
In vivo models provide advantages to study the progression of disease and to identify potential biomarkers to detect and monitor infections. For the human fungal pathogen Cryptococcus neoformans, murine intranasal models aim to recapitulate natural infection from inhalation of desiccated fungal cells from the environment and permit monitoring of disease over time. In this chapter, we describe the establishment of a murine model for cryptococcosis and the subsequent collection of organs, tissues, and fluids for sampling. These samples may support novel diagnostic strategies and opportunities to monitor dissemination of the fungal cells throughout the host and propose new treatment options to combat disease.
Subject(s)
Cryptococcosis , Cryptococcus neoformans , Disease Models, Animal , Animals , Cryptococcus neoformans/pathogenicity , Cryptococcosis/microbiology , Cryptococcosis/diagnosis , Mice , Specimen Handling/methods , HumansABSTRACT
One of the standard assays for the fungal pathogen Cryptococcus neoformans is the glucuronoxylomannan (GXM) ELISA. This assay utilizes monoclonal antibodies targeted against the critical virulence factor, the polysaccharide (PS) capsule. GXM ELISA is one of the most used assays in the field used for diagnosis of cryptococcal infection, quantification of PS content, and determination of binding specificity for antibodies. Here we present three variations of the GXM ELISA used by our group-indirect, capture, and competition ELISAs. We have also provided some history, perspective, and notes on these methods, which we hope will help the reader choose, and implement, the best assay for their research.While it has long been referred to as the GXM ELISA, we also suggest a name update to better reflect our updated understanding of the polysaccharide antigens targeted by this assay. The Cryptococcal PS ELISA is a more accurate description of this set of methodologies and the antigens they measure. Finally, we discuss the limitations of this assay and put forth future plans for expanding the antigens assayed by ELISA.
Subject(s)
Cryptococcosis , Cryptococcus neoformans , Enzyme-Linked Immunosorbent Assay , Polysaccharides , Enzyme-Linked Immunosorbent Assay/methods , Cryptococcus neoformans/immunology , Cryptococcosis/diagnosis , Cryptococcosis/microbiology , Cryptococcosis/immunology , Polysaccharides/analysis , Polysaccharides/immunology , Humans , Antigens, Fungal/immunology , Antigens, Fungal/analysis , Fungal Polysaccharides/immunology , Fungal Polysaccharides/analysis , Antibodies, Monoclonal/immunology , Antibodies, Fungal/immunologyABSTRACT
The importance of humoral immunity to fungal infections remains to be elucidated. In cryptococcosis, patients that fail to generate antibodies against antigens of the fungus Cryptococcus neoformans are more susceptible to the disease, demonstrating the importance of these molecules to the antifungal immune response. Historically, antibodies against C. neoformans have been applied in diagnosis, therapeutics, and as important research tools to elucidate fungal biology. Throughout the process of generating monoclonal antibodies (mAbs) from a single B-cell clone and targeting a single epitope, several immunization steps might be required for the detection of responsive antibodies to the antigen of interest in the serum. This complex mixture of antibodies comprises the polyclonal antibodies. To obtain mAbs, B-lymphocytes are harvested (from spleen or peripheral blood) and fused with tumor myeloma cells, to generate hybridomas that are individually cloned and specifically screened for mAb production. In this chapter, we describe all the necessary steps, from the immunization to polyclonal antibody harvesting, hybridoma generation, and mAb production and purification. Additionally, we discuss new cutting-edge approaches for generating interspecies mAbs, such as humanized mAbs, or for similar species in distinct host backgrounds.
Subject(s)
Antibodies, Fungal , Antibodies, Monoclonal , Cryptococcus neoformans , Hybridomas , Cryptococcus neoformans/immunology , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/isolation & purification , Animals , Humans , Hybridomas/immunology , Antibodies, Fungal/immunology , Antibodies, Fungal/isolation & purification , Mice , B-Lymphocytes/immunology , Cryptococcosis/immunology , Cryptococcosis/diagnosis , Antigens, Fungal/immunology , ImmunizationSubject(s)
Cryptococcus , Meningitis, Cryptococcal , Humans , Antifungal Agents/therapeutic use , Cryptococcosis/diagnosis , Cryptococcosis/epidemiology , Cryptococcosis/immunology , Cryptococcosis/therapy , Cryptococcus/immunology , Cryptococcus/pathogenicity , Immunocompromised Host , Meningitis, Cryptococcal/diagnosis , Meningitis, Cryptococcal/epidemiology , Meningitis, Cryptococcal/immunology , Meningitis, Cryptococcal/therapy , HIV Infections/drug therapy , HIV Infections/immunology , Host-Pathogen Interactions/immunology , Risk Factors , Immunosuppressive Agents/adverse effects , Antigens, Fungal/blood , Antigens, Fungal/cerebrospinal fluid , Intracranial Hypertension/etiology , Intracranial Hypertension/therapy , Immune Reconstitution Inflammatory Syndrome/etiologyABSTRACT
Cryptococcus is an encapsulated yeast that is found in both yeast and hyphal forms. Cryptococcus neoformans and C. gattii are the most medically important species, causing disease in both immunocompromised and immunocompetent individuals. A large, friable, sublingual mass was surgically resected from the oral cavity of a pink-tongued skink (Cyclodomorphus gerarrdii). Histopathology, fungal culture and PCR testing with Sanger sequencing confirmed granulomatous inflammation containing large numbers of yeasts identified as Cryptococcus neoformans var. grubii. Surgical excision and treatment with oral amphotericin B and terbinafine was unsuccessful in managing the infection. On Day 67 after surgery, Cryptococcus DNA was detected by PCR in the blood but not oral, cloacal or skin swabs. The skink was euthanised 72 days after surgery due to anorexia, weight loss and progressive neurological signs of disease. Necropsy results showed disseminated cryptococcosis, including meningoencephalitis and ventriculitis. Two in-contact pink-tongued skinks remained asymptomatic and PCR-negative during 198 days of observation. This case suggests Cryptococcus infections should be considered for oral masses presenting with or without neurological signs in skinks. Further investigation is required to determine the best treatment options for disseminated cryptococcosis in reptiles. This report describes the third reported case of Cryptococcus in a reptile and the first case of cryptococcosis in a pink-tongued skink. It is also the first report of Cryptococcus in a reptile identified to the variety level using PCR, including in whole blood samples.
Subject(s)
Cryptococcosis , Cryptococcus neoformans , Lizards , Animals , Cryptococcosis/veterinary , Cryptococcosis/diagnosis , Cryptococcosis/drug therapy , Cryptococcosis/microbiology , Cryptococcus neoformans/isolation & purification , Lizards/microbiology , Antifungal Agents/therapeutic use , Polymerase Chain Reaction/veterinary , Fatal Outcome , Male , Amphotericin B/therapeutic useABSTRACT
Cryptococcosis, a globally distributed mycotic disease caused by Cryptococcus neoformans or C. gattii, has been extensively studied in various domestic animals and humans. However, non-domestic species have often been overlooked in the literature, with limited attention given to their susceptibility and contribution to the epidemiology of the disease. In this study, a captive two-year-old Cape hyrax in a Japanese zoo exhibited neurological symptoms and torticollis, ultimately succumbing to the infection. Necropsy and pathological analyses, including histopathological techniques and PCR, revealed the presence of C. neoformans in the lungs, cerebrum, and internal auditory canal. While cryptococcosis has been reported in various wild animals globally, this case represents the first documented cryptococcosis in Cape hyrax.
Subject(s)
Animals, Zoo , Cryptococcosis , Cryptococcus neoformans , Animals , Cryptococcosis/veterinary , Cryptococcosis/pathology , Cryptococcosis/microbiology , Cryptococcosis/diagnosis , Cryptococcus neoformans/isolation & purification , Fatal Outcome , Male , Hyraxes , Lung/pathology , Lung/microbiology , Cerebrum/pathology , Cerebrum/microbiologyABSTRACT
Although cryptococcosis is the most common systemic fungal disease of cats, abdominal involvement is rarely reported. The pathogenesis of cryptococcosis usually involves sinonasal colonisation, followed by tissue invasion and sinonasal infection, with possible subsequent spread to the lungs and/or direct extension into the central nervous system (CNS), for example, via the cribriform plate. Further haematogenous spread can occur to any tissue, including skin and the CNS. This report describes a case of disseminated cryptococcosis due to Cryptococcus neoformans species complex in a 13-year-old cat, the fourth documented Australian feline case with abdominal involvement. The cat presented with a chronic history of upper respiratory disease that progressed to severe lethargy and anorexia. An autopsy revealed striking peritonitis with multifocal abdominal involvement affecting the liver, spleen, adrenal glands, kidneys, pancreas and mesentery. Cryptococcal organisms were also observed in organs within the thoracic cavity, sinonasal tissues and the CNS. Testing of abdominal fluid and serum for cryptococcal antigen using a commercially available lateral flow assay using neat fluid specimen initially tested false-negative. However, after dilution of the sample to 1:64, a positive result was obtained, confirming a postzone phenomenon. Taken together, the collective findings were indicative of widely disseminated cryptococcosis due to Cryptococcus neoformans with atypical involvement of the abdominal cavity.
Subject(s)
Cat Diseases , Cryptococcosis , Cryptococcus neoformans , Animals , Cryptococcosis/veterinary , Cryptococcosis/diagnosis , Cryptococcus neoformans/isolation & purification , Cats , Cat Diseases/microbiology , Cat Diseases/diagnosis , Male , Antigens, Fungal , Fatal Outcome , False Negative ReactionsABSTRACT
BACKGROUND: Cryptococcal osteomyelitis is a rare and potentially serious condition, typically encountered in individuals with compromised immune systems. This case underscores the unusual occurrence of disseminated Cryptococcosis in an immunocompetent person, involving multiple bones and lungs, with Cryptococcus neoformans identified as the causative agent. CASE PRESENTATION: An Indonesian man, previously in good health, presented with a chief complaint of successive multiple bone pain lasting for more one month, without any prior history of trauma. Additionally, he reported a recent onset of fever. On physical examination, tenderness was observed in the left lateral chest wall and right iliac crest. Laboratory findings indicated mildly elevated inflammatory markers. A computed tomography (CT) scan of the chest revealed an ovoid solid nodule in the right lower lung and multifocal osteolytic lesions in the sternum, ribs, and humeral head. A magnetic resonance imaging (MRI) study of the sacrum showed multiple lesions in the bilateral iliac bone and the lower L4 vertebral body. Confirmation of Cryptococcal osteomyelitis involved a fine-needle biopsy and culture, identifying Cryptococcus neoformans in the aspirate. The patient responded positively to targeted antifungal treatments, leading to a gradual improvement in his condition. CONCLUSIONS: This case emphasizes the need to consider Cryptococcus neoformans osteomyelitis in immunocompetent patients with bone pain. A definitive diagnosis involves a fine-needle biopsy for pathology and culture, and prompt initiation of appropriate antifungal treatment has proven effective in preventing mortality.