Invasive Mold Infections Research Articles

Opportunistic Invasive Fungal Infections Mimicking Progression of Non–Small-Cell Lung Cancer

BackgroundMany studies have shown that invasive pulmonary aspergillosis, cryptococcosis, and mucormycosis can mimic radiographic and clinical features of primary lung cancer. However, more research surveying the incidence and outcomes of these fungal infections among patients with a history of lung cancer is needed. The aim of this study was to describe the occurrence and clinical outcomes of opportunistic invasive fungal infections that can mimic tumors in non–small-cell lung cancer patients. Patients and MethodsPatients seen at Stanford University Medical Center from January 1, 2007, to May 1, 2020, with pulmonary aspergillosis, cryptococcosis, or mucormycosis after non–small-cell lung cancer (NSCLC) diagnosis were reviewed. The European Organization for Research and Treatment of Cancer National Institute of Allergy and Infectious Diseases Mycoses Study Group criteria was used to classify patients with evidence of proven or probable invasive fungal infection within our cohort. ResultsA total of 12 patients with proven or probable invasive mold infection (including 8 cases of aspergillosis) and 1 patient with proven cryptococcosis were identified, without any cases of mucormycosis. Of this cohort, 6 patients (46%) showed radiographic findings that were found to be most consistent with lung cancer by radiologists. Eight cases (62%) were suspected of cancer recurrence or progression by the treatment team on the basis of additional considerations of medical history and clinical symptoms. Most patients had active NSCLC or had a history of recurrence without active NSCLC at the time of fungal discovery (11 patients; 85%). Most patients died without full recovery (7 patients; 54%). ConclusionsInvasive pulmonary aspergillosis and cryptococcosis can often be mistaken as cancer recurrence or progression in patients with a history of NSCLC because of mimicking radiographic and clinical characteristics.

Diagnosis of Breakthrough Fungal Infections in the Clinical Mycology Laboratory: An ECMM Consensus Statement.

Breakthrough invasive fungal infections (bIFI) cause significant morbidity and mortality. Their diagnosis can be challenging due to reduced sensitivity to conventional culture techniques, serologic tests, and PCR-based assays in patients undergoing antifungal therapy, and their diagnosis can be delayed contributing to poor patient outcomes. In this review, we provide consensus recommendations on behalf of the European Confederation for Medical Mycology (ECMM) for the diagnosis of bIFI caused by invasive yeasts, molds, and endemic mycoses, to guide diagnostic efforts in patients receiving antifungals and support the design of future clinical trials in the field of clinical mycology. The cornerstone of lab-based diagnosis of breakthrough infections for yeast and endemic mycoses remain conventional culture, to accurately identify the causative pathogen and allow for antifungal susceptibility testing. The impact of non-culture-based methods are not well-studied for the definite diagnosis of breakthrough invasive yeast infections. Non-culture-based methods have an important role for the diagnosis of breakthrough invasive mold infections, in particular invasive aspergillosis, and a combination of testing involving conventional culture, antigen-based assays, and PCR-based assays should be considered. Multiple diagnostic modalities, including histopathology, culture, antibody, and/or antigen tests and occasionally PCR-based assays may be required to diagnose breakthrough endemic mycoses. A need exists for diagnostic tests that are effective, simple, cheap, and rapid to enable the diagnosis of bIFI in patients taking antifungals.

THE TALE OF A HALO SIGN

SESSION TITLE: Fellows Imaging Posters SESSION TYPE: Fellow Case Report Posters PRESENTED ON: October 18-21, 2020 INTRODUCTION: Halo sign on chest imaging is characterized by a circular area of ground-glass attenuation around a central pulmonary nodule or mass. It is most commonly associated with invasive fungal infections; however, can also be seen in a wide range of neoplastic and inflammatory conditions. Here we present a case of angio-invasive neoplasm presenting with characteristic halo sign in an immunocompromised patient. CASE PRESENTATION: A 62-year-old male with alcoholic cirrhosis post orthotopic liver transplant on chronic immunosuppression with recurrent cirrhosis of his graft liver was initially admitted for management of acute hepatic encephalopathy. A CT scan of his chest showed numerous part solid centrilobular nodules bilaterally with central solid component and surrounding ground glass halo. Invasive fungal infection was suspected, and micafungin was initiated. Serum aspergillus antigen and beta D-glucan were negative. He underwent bronchoscopy with bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB). BAL was negative for bacterial, fungal and acid-fast bacilli cultures. Pathology from the TBB showed atypical epithelioid cells that could be reactive or due to a metastatic disease. A repeat CT scan performed after 10 days of antifungal therapy showed a significant increase in size of the previously noted centrilobular nodules with surrounding ground glass halo. Surgical open lung biopsy was considered but he was deemed to be a poor surgical candidate due to his frailty and comorbidities. Transbronchial cryobiopsy was then performed which showed poorly differentiated epithelioid neoplasm predominantly within the vascular spaces. Based on immunohistochemical staining positive for both cytokeratin and vascular markers, a differential diagnosis of epithelioid angiosarcoma vs. epithelioid sarcoma was made. After detailed discussion with him and his family in regards to his overall prognosis, he wished for no further testing or interventions and chose to go home with hospice. DISCUSSION: Halo sign indicates presence of hemorrhagic nodules. It was first described in immunocompromised patients with invasive pulmonary aspergillosis where the nodule itself is a focus of pulmonary infarction with surrounding alveolar hemorrhage. However, it has now been associated with a variety of pulmonary diseases including both infectious and non-infectious hemorrhagic nodules, tumor cell infiltration and few non-hemorrhagic etiologies. Metastatic lung disease from hyper vascular tumors such as angiosarcoma, chondrosarcoma and melanoma can form a halo of ground glass attenuation as a result of peritumoral hemorrhage or infiltration. Malignancy is a leading cause of death in post liver transplant patients and should always be considered in the evaluation of these patients. CONCLUSIONS: While evaluating halo sign a wide differential should be considered including malignancy in a post liver transplant patient. Reference #1: Georgiadou SP, Sipsas NV, Marom EM, Kontoyiannis DP. The diagnostic value of halo and reversed halo signs for invasive mold infections in compromised hosts. Clin Infect Dis. 2011;52(9):1144-1155. doi:10.1093/cid/cir122 Reference #2: Parrón M, Torres I, Pardo M, Morales C, Navarro M, Martínez-Schmizcraft M. Signo del halo en la tomografía computarizada de tórax: diagnóstico diferencial con correlación anatomopatológica [The halo sign in computed tomography images: differential diagnosis and correlation with pathology findings]. Arch Bronconeumol. 2008;44(7):386-392. DISCLOSURES: No relevant relationships by Sadia Benzaquen, source=Web Response No relevant relationships by Ena Gupta, source=Web Response No relevant relationships by ATUL MATTA, source=Web Response No relevant relationships by Corrado Minimo, source=Web Response No relevant relationships by Carlos Oberto, source=Web Response

Future challenges and chances in the diagnosis and management of invasive mould infections in cancer patients

Diagnosis, treatment, and management of invasive mould infections (IMI) are challenged by several risk factors, including local epidemiological characteristics, the emergence of fungal resistance and the innate resistance of emerging pathogens, the use of new immunosuppressants, as well as off-target effects of new oncological drugs. The presence of specific host genetic variants and the patient's immune system status may also influence the establishment of an IMI and the outcome of its therapy. Immunological components can thus be expected to play a pivotal role not only in the risk assessment and diagnosis, but also in the treatment of IMI. Cytokines could improve the reliability of an invasive aspergillosis diagnosis by serving as biomarkers as do serological and molecular assays, since they can be easily measured, and the turnaround time is short. The use of immunological markers in the assessment of treatment response could be helpful to reduce overtreatment in high risk patients and allow prompt escalation of antifungal treatment. Mould-active prophylaxis could be better targeted to individual host needs, leading to a targeted prophylaxis in patients with known immunological profiles associated with high susceptibility for IMI, in particular invasive aspergillosis. The alteration of cellular antifungal immune response through oncological drugs and immunosuppressants heavily influences the outcome and may be even more important than the choice of the antifungal treatment. There is a need for the development of new antifungal strategies, including individualized approaches for prevention and treatment of IMI that consider genetic traits of the patients.Lay Anticancer and immunosuppressive drugs may alter the ability of the immune system to fight invasive mould infections and may be more important than the choice of the antifungal treatment. Individualized approaches for prevention and treatment of invasive mold infections are needed.

Pentraxin-3 polymorphisms and pulmonary fungal disease in non-neutropenic patients.

BackgroundPentraxin 3 (PTX3) plays a non-redundant role in innate immunity against fungal diseases. Although single nucleotide polymorphisms (SNPs) of PTX3 are associated with a higher risk of invasive aspergillosis among the immunosuppressed population and chronic obstructive pulmonary disease patients, it is unknown whether PTX3 genetic variants influence the risk of pulmonary fungal disease in immunocompetent patients.MethodsTo investigate the association between PTX3 gene polymorphisms and pulmonary mycosis in non-neutropenic patients, we conducted a case-control study in a tertiary hospital department. Forty-five patients were identified using the criteria of the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC-MSG) and enrolled in the case group. Of these patients, 15 had allergic bronchopulmonary aspergillosis (ABPA), 10 had invasive pulmonary aspergillosis (IPA), 18 had pulmonary cryptococcosis, and 2 had other types of pulmonary mycosis. One hundred and twenty-two non-neutropenic inpatients not infected by fungal disease were randomly selected as the control group. We detected three SNPs (rs2305619, rs3816527, and rs1840680) within the PTX3 gene using polymerase chain reaction sequencing and compared their associations with different types of pulmonary fungal disease.ResultsThree SNPs were consistent with Hardy-Weinberg equilibrium (HWE). SNP rs2305619 was in linkage disequilibrium with rs3816527 (D’=0.85) and rs1840680 (D’=0.85), respectively. There was no difference in the genotypic distribution and haplotype frequency of the SNPs between the case group and the control group. When we focused on invasive mold infections as a subgroup, we found that the SNP rs3816527 CC homozygote was associated with a higher risk of IPA (OR, 7.37; 95% CI, 0.93–44.44; P=0.033), while the rs3816527 AA homozygote might lower the risk of pulmonary cryptococcosis (OR, 0.35; 95% CI, 0.11–0.96; P=0.047). No genotypic distribution differences were observed for the other two SNPs (rs2305619 and rs1840680). When it came to the comparison between ABPA subgroup and control group, no difference in single nucleotide polymorphism was observed.ConclusionsThis study showed that the SNP rs3816527 is associated with IPA in non-neutropenic patients. Further investigations in large populations are needed to validate this genetic predisposition. Functional studies are also required.

SCEDOSPORIUM APIOSPERMUM AND LOMENTOSPORA PROLIFICANS IN LUNG TRANSPLANT PATIENTS: A SINGLE CENTRE EXPERIENCE OVER 24 YEARS

Introduction: Sceodosporium and Lomentospora (Sceodosporium/Lomentospora) species cause multi resistant and life-threatening respiratory tract infections. Amongst lung transplant recipients, these emerging pathogens now represent the second most common cause of invasive mould infections following Aspergillus.(1,2) The current epidemiology, diagnostic algorithm and management is unknown. Methods: A retrospective single centre audit of all sputum and bronchoscopy samples positive for Sceodosporium/Lomentospora species in lung transplant patients over a 15-year period (1995-2019). Patients were defined as either being colonised with the pathogen or diagnosed with invasive fungal disease (IFD) as per 2010 the International Society of Heart and Lung Transplant (ISHLT) definitions(3). Results: Since 1995, 30 from a total lung transplant cohort of 959 patients (3.13%) cultured Scedosporium/Lomentospora from either sputum sample or bronchoscopic washings. The first isolate was detected in 2014 (n=9), with cases detected yearly thereafter (2015 [n=1]; 2016 [n=8]; 2017 [n=6]; n=2018 [n=6]). Nineteen (63%) Scedosporium/Lomentospora isolation was classified as invasive (IFD), and 27% (n=11) were deemed as colonisers. The median time to first culture from lung transplant was 929 days [95% CI 1089-3168 days]. The majority of patients (n = 19; 63.3%) had received an antifungal agent prior to the first positive culture (posaconazole [n=15; 79%], voriconazole [n=2;11%], posaconazole + terbinadine [n=1; 5%], terbinafine [n=1; 5%]). The most common antifungal agent used for treatment was posaconazole (n=16) followed by terbinafine (n=9), and multiple sequential agents were used in 8 (26.7%). Median duration of therapy was 364 days [95% CI 264-748 days]. Median FEV1 increased by 400mls following six months treatment of Scedosporium/Lomentospora.Overall survival amongst patients with Scedosporium/Lomentospora detected from respiratory sampling superior compared to the remainder of the LTx cohort (65% vs 34% p=0.04)Discussion: Our experience is in keeping with previous smaller studies, however this sample size and duration of follow up for Scedosporum/Lomentospora has not previously been reported. Our single centre 24-year experience suggests that the incidence of Scedosporium/Lomentospora in respiratory samples of lung transplant recipients is emerging. Most patients with positive Scedosporum/Lomentospora on sputum/bronchoscopic washings have had prior exposure to anti-fungal agents. Conclusion: Treatment of Scedosporium/Lomentospora infection results in improved lung function, and in good overall survival rates. Further surveillance is required to fully elucidate the impact of these organisms on lung transplant recipients.

Review of the Novel Investigational Antifungal Olorofim.

The incidence of invasive fungal infections caused by molds and endemic fungi is increasing. There is also concern regarding increased rates of reduced susceptibility or frank resistance among Aspergillus and Coccidioides species, while Scedosporium species, Lomentospora prolificans, and Fusarium species are inherently less susceptible or intrinsically resistant to clinically available antifungals. Olorofim (formerly F901318) is the first member of the orotomide class of antifungals to be evaluated clinically for the treatment of invasive mold infections. This agent inhibits dihydroorotate dehydrogenase, a key enzyme in the biosynthesis of pyrimidines. Olorofim has activity against many molds and thermally dimorphic fungi, including species that are resistant to azoles and amphotericin B, but lacks activity against yeasts and the Mucorales. It is currently being developed for both oral and intravenous administration. Although published clinical outcome data have been limited to case reports to date, the results against invasive and refractory infections are promising. This review describes the mechanism of action of olorofim, its in vitro spectrum of activity, and what is currently known about its pharmacokinetic profile and clinical efficacy.

Association between the kinetics of cytomegalovirus reactivation evaluated in terms of the area under the curve of cytomegalovirus antigenemia and invasive mold infection during the post-engraftment phase after allogeneic hematopoietic stem cell transplantation.

We evaluated the clinical impact of cytomegalovirus (CMV) reactivation calculated in terms of the area under the curve of CMV antigenemia (CMV-AUC) on the development of invasive mold infection (IMI) in the post-engraftment phase after allogeneic hematopoietic stem cell transplantation (HSCT). Among 394 consecutive patients who underwent their first allogeneic HSCT at our center between 2007 and 2018, 335 were included after excluding patients with a past history of invasive fungal disease (IFD), the development of IFD before engraftment, engraftment failure, or early death within 30days. CMV antigenemia (CMV-AG) was monitored weekly after engraftment and 3 or more cells/2 slides were regarded as positive. CMV-AUC was calculated by the trapezoidal method using the number of CMV-AG after logarithmic transformation and the duration in weeks and was added until negative conversion. Patients with CMV reactivation were divided into low and high CMV-AUC groups using the median value of CMV-AUC as a threshold. There were 17 proven/probable IMIs including one mucormycosis and 16 probable invasive aspergillosis, and the 2-year cumulative incidence was 1.0% in the negative CMV-AUC group (n=136), 3.3% in the low CMV-AUC group (n=98) and 13.8% in the high CMV-AUC group (n=101) (P=.001). In a multivariate analysis, grade II-IV acute GVHD (HR 3.74) and CMV-AUC (HR low 1.25, high 5.91) were identified as independent significant factors associated with a higher incidence of IMI. Cytomegalovirus kinetics evaluated in terms of CMV-AUC were significantly associated with the development of IMI in the post-engraftment phase after allogeneic HSCT.

In Vitro Evaluation of Radiolabeled Amphotericin B for Molecular Imaging of Mold Infections.

Invasive pulmonary aspergillosis and mucormycosis are life-threatening complications in immunocompromised patients. A rapid diagnosis followed by early antifungal treatment is essential for patient survival. Given the limited spectrum of biomarkers for invasive mold infections, recent studies have proposed the use of radiolabeled siderophores or antibodies as molecular probes to increase the specificity of radiological findings by nuclear imaging modalities. While holding enormous diagnostic potential, most of the currently available molecular probes are tailored to the detection of Aspergillus species, and their cost-intensive and sophisticated implementation restricts their accessibility at less specialized centers. In order to develop cost-efficient and broadly applicable tracers for pulmonary mold infections, this study established streamlined and high-yielding protocols to radiolabel amphotericin B (AMB) with the gamma emitter technetium-99m (99mTc-AMB) and the positron emitter gallium-68 (68Ga-AMB). The radiochemical purity of the resulting tracers consistently exceeded 99%, and both probes displayed excellent stability in human serum (>98% after 60 to 240 min at 37°C). The uptake kinetics by representative mold pathogens were assessed in an in vitro Transwell assay using infected endothelial cell layers. Both tracers accumulated intensively and specifically in Transwell inserts infected with Aspergillus fumigatus, Rhizopus arrhizus, and other clinically relevant mold pathogens compared with their accumulation in uninfected inserts and inserts infected with bacterial controls. Inoculum-dependent enrichment was confirmed by gamma counting and autoradiographic imaging. Taken together, this pilot in vitro study proposes 99mTc-AMB and 68Ga-AMB to be facile, stable, and specific probes, meriting further preclinical in vivo evaluation of radiolabeled amphotericin B for molecular imaging in invasive mycoses.

Isavuconazole Is as Effective as and Better Tolerated Than Voriconazole for Antifungal Prophylaxis in Lung Transplant Recipients

Invasive fungal infections (IFIs) are common following lung transplantation. Isavuconazole is unstudied as prophylaxis in organ transplant recipients. We compared effectiveness and tolerability of isavuconazole and voriconazole prophylaxis in lung transplant recipients. A single-center, retrospective study of patients who received isavuconazole (September 2015-February 2018) or voriconazole (September 2013-September 2015) for antifungal prophylaxis. IFIs were defined by EORTC/MSG criteria. Patients received isavuconazole (n = 144) or voriconazole (n = 156) for median 3.4 and 3.1 months, respectively. Adjunctive inhaled amphotericin B (iAmB) was administered to 100% and 41% of patients in the respective groups. At 1 year, 8% of patients receiving isavuconazole or voriconazole developed IFIs. For both groups, 70% and 30% of IFIs were caused by molds and yeasts, respectively, and breakthrough IFI (bIFI) rate was 3%. Outcomes did not significantly differ for patients receiving or not receiving iAmB. Independent risk factors for bIFI and breakthrough invasive mold infection (bIMI) were mold-positive respiratory culture and red blood cell transfusion >7 units at transplant. Bronchial necrosis >2 cm from anastomosis and basiliximab induction were also independent risk factors for bIMI. Isavuconazole and voriconazole were discontinued prematurely due to adverse events in 11% and 36% of patients, respectively (P = .0001). Most common causes of voriconazole and isavuconazole discontinuation were hepatotoxicity and lack of oral intake, respectively. Patients receiving ≥90 days prophylaxis had fewer IFIs at 1 year (3% vs 9%, P = .02). IFIs were associated with increased mortality (P = .0001) and longer hospitalizations (P = .0005). Isavuconazole was effective and well tolerated as antifungal prophylaxis following lung transplantation.

Antifungal prophylaxis during 7 + 3 induction chemotherapy for acute myeloid leukemia is associated with improved survival, in a setting with low incidence of invasive mold infections.

Invasive fungal infections (IFIs) are a major cause of morbidity and mortality in patients undergoing induction chemotherapy for acute myeloid leukemia (AML). In this patient population, antifungal prophylaxis (AP) has been associated with decreased incidence of IFIs and better survival. However, some centers have not adopted AP during induction chemotherapy for AML, as it is unclear whether AP improves outcomes in settings where the incidence of invasive mold infections is low. We retrospectively assessed the differences in clinical outcomes and resource utilization in patients undergoing 7 + 3 induction chemotherapy for AML, after implementing a policy of AP as part of a dedicated inpatient malignant hematology service (HS) at Rhode Island Hospital. Between January 1, 2007 and April 1, 2019, 56 patients with AML received AP during 7 + 3 induction chemotherapy and 52 patients did not, without significant differences in their baseline characteristics. Use of AP was associated with less proven or probable IFI (0% vs. 6%, P = 0.1) and lower all-cause in-hospital mortality (7% vs. 21%, P < 0.05), without significant increases in resource utilization or toxicities. Empiric and targeted antifungal therapies were more frequently started in the non-AP group (69%) than changed in the AP group (41%, P < 0.005). Having a dedicated inpatient malignant hematology service was also associated with improved outcomes. However, use of AP was associated with better survival (30-day post-induction survival log-rank P < 0.05), prior to the implementation of this clinical service as well, which is suggestive of an independent benefit from AP.

Low-dose liposomal amphotericin B for antifungal prophylaxis in paediatric allogeneic haematopoietic stem cell transplantation.

Primary antifungal prophylaxis in paediatric allogeneic HSCT recipients is mainly based on azoles, which can have related toxicity and drug interactions. Low-dose liposomal amphotericin B (L-AmB) is an attractive intravenous alternative because of its low toxicity and lower risk of interactions. To evaluate the effectiveness and safety of L-AmB (1 mg/kg/day) for primary antifungal prophylaxis in pre-engraftment paediatric HSCT patients. Retrospective, observational study including all consecutive patients aged ≤18 years who underwent HSCT and received antifungal prophylaxis with intravenous L-AmB (1 mg/kg/day, from day -1 to 48 h before discharge) between January 2012 and December 2016. In total, 125 HSCT procedures in 118 patients were included, median age 7.2 years (IQR 4.2-11.5). Haematological malignancies were the main underlying condition (63.6%), and 109 (87.2%) were considered at high risk for invasive fungal infection (IFI). Ten patients (7.7%), all high risk, developed breakthrough IFI (three Candida spp., seven invasive mould infections) and tended to have higher overall mortality. The only statistically significant risk factor for IFI was cytomegalovirus co-infection. Adverse events, all grade I, occurred in 25 (20%), requiring L-AmB withdrawal in one case. Overall survival at 30 days was 99.2%. At study completion, one patient had died of IFI. The incidence of breakthrough IFI was comparable to that of previous reports, with a very low rate of significant toxicity. Thus, prophylactic L-AmB may be a safe, effective option for antifungal prophylaxis in the pre-engraftment phase for children undergoing HSCT, even those at high risk.

Pharmacokinetics and Pharmacodynamics of Posaconazole.

Posaconazole is typically used for preventing invasive yeast and mold infections such as invasive aspergillosis in high-risk immunocompromised patients. The oral suspension was the first released formulation and many pharmacokinetic and pharmacodynamic studies of this formulation have been published. Erratic absorption profiles associated with this formulation were widely reported. Posaconazole exposure was found to be significantly influenced by food and many gastrointestinal conditions, including pH and motility. As a result, low posaconazole plasma concentrations were obtained in large groups of patients. These issues of erratic absorption urged the development of the subsequently marketed delayed-release tablet, which proved to be associated with higher and more stable exposure profiles. Shortly thereafter, an intravenous formulation was released for patients who are not able to take oral formulations. Both new formulations require a loading dose on day 1 to achieve high posaconazole concentrations more quickly, which was not possible with the oral suspension. So far, there appears to be no evidence of increased toxicity correlated to the higher posaconazole exposure achieved with the regimen for these formulations. The higher systemic availability of posaconazole for the delayed-release tablet and intravenous formulation have resulted in these two formulations being preferable for both prophylaxis and treatment of invasive fungal disease. This review aimed to integrate the current knowledge on posaconazole pharmacokinetics, pharmacodynamics, major toxicity, existing resistance, clinical experience in special populations, and new therapeutic strategies in order to get a clear understanding of the clinical use of this drug.

Impact of immunosuppressive and antifungal drugs on PBMC- and whole blood-based flow cytometric CD154+ Aspergillus fumigatus specific T-cell quantification.

Flow cytometric quantification of CD154+ mould specific T-cells in antigen-stimulated peripheral blood mononuclear cells (PBMCs) or whole blood has been described as a supportive biomarker to diagnose invasive mould infections and to monitor therapeutic outcomes. As patients at risk frequently receive immunosuppressive and antifungal medication, this study compared the matrix-dependent impact of representative drugs on CD154+ T-cell detection rates. PBMCs and whole blood samples from healthy adults were pre-treated with therapeutic concentrations of liposomal amphotericin B, voriconazole, posaconazole, cyclosporine A (CsA) or prednisolone. Samples were then stimulated with an Aspergillus fumigatus lysate or a viral antigen cocktail (CPI) and assessed for CD154+ T-helper cell frequencies. Specific T-cell detection rates and technical assay properties remained largely unaffected by exposure of both matrices to the studied antifungals. By contrast, CsA and prednisolone pre-treatment of isolated PBMCs and whole blood adversely impacted specific T-cell detection rates and caused elevated inter-replicate variation. Unexpectedly, the whole blood-based protocol that uses additional α-CD49d co-stimulation was less susceptible to CsA and prednisolone despite prolonged drug exposure in the test tube. Accordingly, addition of α-CD49d during PBMC stimulation partially attenuated the impact of immunosuppressive drugs on test performance. Translating these results into the clinical setting, false-negative results of CD154+ antigen-specific T-cell quantification need to be considered in patients receiving T-cell-active immunosuppressive medication. Optimized co-stimulation regimes with α-CD49d could contribute to an improved feasibility of functional T-cell assays in immunocompromised patient populations.

Evaluation of Universal versus Preemptive Antifungal Prophylaxis in Lung Transplantation

Infections from various fungi are increasingly recognized in the setting of lung transplant and guidelines do not distinguish one superior strategy between universal or preemptive. Limited studies exist comparing the two approaches and this study aims to evaluate the incidence mold infections post lung transplant (LTx) before and after the implementation of a protocol change. Our institution's PRE-protocol (PRE) included a pre-emptive prophylaxis for high risk IMI (invasive mold infection) with positive donor or recipient mold colonizations. POST-protocol implementation, a universal prophylaxis approach using itraconazole 200mg PO BID for lifetime post-transplant was employed. A single-center, retrospective chart review was conducted on adult LTx recipients. Patients receiving LTx between 1/2012-5/2016 were included in the PRE cohort and those between 8/2016-5/2019 were included in the POST cohort. A total of 150 patients were included in the PRE cohort and 127 in the POST cohort. Baseline demographics were similar among the cohorts in regards to mean age, gender, bilateral transplant and race. Incidence of IMI or disseminated fungal disease was n=13 (9%) vs n=1 (1%) in the PRE vs POST cohort; p=0.002. A total of 94/150 (63%) vs 51/127 (40%); p=.001 patients developed post LTx fungal colonization. The distribution of types of mold (Table 1) show that there was significantly less BAL colonization with aspergillus with the universal prophylaxis protocol (POST) 33 vs 64%. Median time to first fungal colonization was similar in both groups 108 vs 109 days in the respective PRE and POST groups. Discontinuation rates of itraconazole was n=28 (22%) among the POST cohort. While fungal colonization is ubiquitous, invasive and disseminated fungal infections can result in detrimental outcomes post lung transplant. The strategy of lifelong universal prophylaxis for molds may confer decreased colonization rates and in turn lower incidence if invasive and disseminated disease.

Fungal Prophylaxis and Chronic Lung Allograft Dysfunction

Prior studies demonstrate that lung transplant recipients (LTRs) experience the highest rate of invasive mold infections amongst all transplant recipients. Aspergillosis is the most common offender, and aspergillus colonization and invasive disease have been linked to chronic lung allograft dysfunction (CLAD). There are no standardized guidelines regarding fungal prophylaxis strategy or agent selection. This study examined the relationship between post-transplant antifungal prophylaxis and freedom from CLAD. This single-center retrospective analysis evaluated 566 adult LTRs from July 2, 2005 to December 28, 2015. Fungal prophylaxis practices evolved during the study period from targeted antifungal prophylaxis (TAP) to universal antifungal prophylaxis (UAP) for the first 6 months post-transplant with either voriconazole or posaconazole. A Kaplan-Meir analysis and Cox regression were performed to estimate freedom from CLAD at 3 years for TAP versus UAP patients. A similar approach was used to examine the relationship between freedom from CLAD and discharge antifungal selection (none, voriconazole, or posaconazole). Freedom from CLAD at 3 years post-transplant in the TAP versus UAP cohorts was 65% and 68%, respectively (P=0.35). The relative risk of CLAD was not significantly reduced in the UAP cohort even when adjusting for pre-transplant disease, transplant type, and induction therapy (hazard ratio 0.83; 95% confidence interval 0.63-1.13; P=0.24). Freedom from CLAD at 3 years post-transplant with patients grouped by discharge antifungal regimen was as follows: no prophylaxis 67%, voriconazole 68%, and posaconazole 70% (P=0.65). There was also no significant difference in risk of CLAD across patients grouped by discharge antifungal regimen following multivariable adjustment. There was no significant difference in freedom from CLAD at 3 years post-transplant for LTRs managed under a TAP versus UAP strategy or for these patients when grouped by discharge antifungal regimen. These results, while unexpected, pique interest in further investigation, particularly elucidating the relationship between prophylaxis and incidence of fungal colonization and invasive disease. Pursuing these avenues of study will advance our understanding of the utility and benefit of fungal prophylaxis in the LTR population.

A Single-Center, Open-Label Trial of Isavuconazole Prophylaxis against Invasive Fungal Infection in Patients Undergoing Allogeneic Hematopoietic Cell Transplantation.

Isavuconazole is a broad-spectrum triazole approved for treatment of invasive fungal infections (IFIs). In this open-label, single-arm study, we evaluated isavuconazole for antifungal prophylaxis after allogeneic hematopoietic cell transplantation (HCT). Adult patients admitted for first HCT received micafungin 150 mg i.v. daily from admission through day +7 (D+7) post-transplantation (±2 days) followed by isavuconazole prophylaxis (i.v./p.o. 372 mg every 8 hours for 6 doses and then 372 mg daily) through maximum D+98 post-HCT. Patients were followed through D+182. The primary endpoint was prophylaxis failure, defined as discontinuation of prophylaxis for proven/probable IFI; systemic antifungal therapy for >14 days for suspected IFI; toxicity leading to discontinuation; or an adverse event. Between June 2017 and October 2018, 99 patients were enrolled in the study, of whom 95 were included in our analysis. The median patient age was 57 years (interquartile range [IQR], 50 to 66 years). Sixty-four (67%) patients received peripheral blood, 17(18%) received bone marrow, and 14 (15%) received a cord blood allograft for acute leukemia (55%), lymphoma (17%), myelodysplastic syndrome (16%), or another hematologic disease (14%). One-third (n=31; 33%) of patients underwent CD34+-selected HCT. Isavuconazole prophylaxis was given for a median of 90 days (IQR, 87 to 91 days). Ten patients (10.7%) met the primary endpoint. Candidemia occurred in 3 patients (3.1%), 1 of whom had grade III skin acute graft-versus-host disease (GVHD). Toxicity leading to discontinuation occurred in 7 patients (7.4%). The most common toxicity was liver function abnormalities in 5 patients, including grade 1 transaminitis in 2 patients and grade 3 hyperbilirubinemia in 3 patients. Four patients (4.2%) had early discontinuation of isavuconazole for reasons not meeting the primary study endpoint. Six patients died during the study period, including 3 during prophylaxis and 3 during follow-up. No deaths were attributed to isavuconazole. The majority (85%) of allogeneic HCT recipients completed isavuconazole prophylaxis according to protocol. The rate of breakthrough candidemia was 3.1%, and there were no invasive mold infections. Our data support the utility of isavuconazole for antifungal prophylaxis after HCT.

An Open-label Study of APX001 for Treatment of Patients With Invasive Mold Infections Caused by Aspergillus Species or Rare Molds

An Open-label Study of APX001 for Treatment of Patients With Invasive Mold Infections Caused by Aspergillus Species or Rare Molds

Infectious Complications of CAR T-Cell Therapy in Non-Hodgkin Lymphoma

Chimeric Antigen Receptor T-Cell (CART) therapy is a promising therapeutic option for patients with relapsed, refractory lymphomas. Both immunosuppression and infections are complications reported in the post-CART setting. The objective of this study was to identify rates of infectious complications and febrile neutropenia at our institution. This retrospective single center study included 38 patients with relapsed, refractory non-Hodgkin lymphoma (NHL), who received CD19 directed CAR T-cell therapy axicabtagene ciloleucel, as standard of care for the treatment relapsed NHL between January, 2018 and October, 2019. Infectious complications were identified and classified as bacterial, viral, and either as non-invasive fungal or invasive fungal. We found that 79% (30/38) of patients experienced an episode of febrile neutropenia and 22 of the 46 identified infections occurred during an episode of neutropenia. Overall, 79% (30/38) of patients had an identified infection of viral, bacterial, or fungal. Sixteen patients (42%) were found to have bacterial infections, eight patients (21%) viral infections, and 17 (45%) fungal infections. Of the forty-six infections identified, 16 were bacterial, 11 viral, and 19 fungal. The most common sites of bacterial infections noted were blood stream infections (37.5%) and pulmonary (25%) sources. The most common viral infections identified were CMV (36%), BK virus (18%), and RSV (18%). The majority of the fungal infections identified were oral candidiasis (89.5%) with only 2 patients developing an invasive mold infection. Overall, we had notable infectious findings including a high incidence of non-invasive candidiasis and occurrences of opportunistic viral infections associated with CAR-T cell therapy and lymphodepletion as compared to the literature. There is a relative paucity of guidelines and literature on supportive cares as it relates to infectious complications. We ultimately conclude that infectious complications are common adverse effects of CAR-T therapy that require more robust investigation and further evidence-based and consensus driven guidelines.

Immune Parameters for Diagnosis and Treatment Monitoring in Invasive Mold Infection.

Infections caused by invasive molds, including Aspergillus spp., can be difficult to diagnose and remain associated with high morbidity and mortality. Thus, early diagnosis and targeted systemic antifungal treatment remains the most important predictive factor for a successful outcome in immunocompromised individuals with invasive mold infections. Diagnosis remains difficult due to low sensitivities of diagnostic tests including culture and other mycological tests for mold pathogens, particularly in patients on mold-active antifungal prophylaxis. As a result, antifungal treatment is rarely targeted and reliable markers for treatment monitoring and outcome prediction are missing. Thus, there is a need for improved markers to diagnose invasive mold infections, monitor response to treatment, and assist in determining when antifungal therapy should be escalated, switched, or can be stopped. This review focuses on the role of immunologic markers and specifically cytokines in diagnosis and treatment monitoring of invasive mold infections.